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- PublicationA-RafA-Raf (v-raf murine sarcoma 3611 viral oncogene homolog) is a serine/threonine protein kinase of the Raf family that comprises A-Raf, B-Raf and C-Raf. Raf kinases are at the apex of the three-tiered Raf-MEK-ERK/MAPK pathway that features over 150 substrates and regulates many fundamental cellular functions, including proliferation, differentiation, transformation, apoptosis and metabolism. The only commonly accepted substrates for all three Raf kinases are MEK1/2, a pair of dual-specificity kinases that have ERK1/2 as substrates. A-Raf is the least studied member of the Raf family. A-Raf seems to be regulated similarly to C-Raf, with binding to activated Ras initiating the growth-factor-induced activation of A-Raf. In addition, A-Raf activity is regulated by phosphorylation, lipid interactions and protein-protein interactions. For instance, binding of the regulatory subunit of casein kinase II, CK2β, was shown to enhance A-Raf kinase activity. However, A-Raf is a poor MEK kinase with barely measurable catalytic activity, suggesting that A-Raf could have functions outside the MAPK cascade. A-Raf binding to mitochondrial membrane proteins suggests a potential role in mitochondrial transport and anti-apoptotic signaling pathways. Furthermore, the association of A-Raf with the pyruvate kinase M2, M2-PK, causing dimerization and inactivation of M2-PK, may link A-Raf signaling with energy metabolism and the Warburg effect in tumor cells. The generation of A-Raf knock-out mice revealed a role in neuronal migration and development. Recently, alternative A-Raf splice forms encoding truncated A-Raf proteins were identified. Owing to their ability to bind and block activated Ras, they function as physiological dominant-negative Ras inhibitors with roles in differentiation and transformation. A-Raf is expressed in most tissues, but expression levels differ dramatically. Elevated levels were reported in a number of malignancies, although no oncogenic mutations have been found.
266 - PublicationAcute hypoxic exposure and prolyl-hydroxylase inhibition improves synaptic transmission recovery time from a subsequent hypoxic insult in rat hippocampus(Elsevier, 2018-12-15)
; ; ; ; In the CNS short episodes of acute hypoxia can result in a decrease in synaptic transmission which may be fully reversible upon re-oxygenation. Stabilization of hypoxia-inducible factor (HIF) by inhibition of prolyl hydroxylase domain (PHD) enzymes has been shown to regulate the cellular response to hypoxia and confer neuroprotection both in vivo and in vitro. Hypoxic preconditioning has become a novel therapeutic target to induce neuroprotection during hypoxic insults. However, there is little understanding of the effects of repeated hypoxic insults or pharmacological PHD inhibition on synaptic signalling. In this study we have assessed the effects of hypoxic exposure and PHD inhibition on synaptic transmission in the rat CA1 hippocampus. Field excitatory postsynaptic potentials (fEPSPs) were elicited by stimulation of the Schaffer collatoral pathway. 30 min hypoxia (gas mixture 95% N2/5% CO2) resulted in a significant and fully reversible decrease in fEPSP slope associated with decreases in partial pressures of tissue oxygen. 15-30 min of hypoxia was sufficient to induce stabilization of HIF in hippocampal slices. Exposure to a second hypoxic insult after 60 min resulted in a similar depression of fEPSP slope but with a significantly greater rate of recovery of the fEPSP. Prior single treatment of slices with the PHD inhibitor, dimethyloxalylglycine (DMOG) also resulted in a significantly greater rate of recovery of fEPSP post hypoxia. These results suggest that hypoxia and ‘pseudohypoxia’ preconditioning may improve the rate of recovery of hippocampal neurons to a subsequent acute hypoxia.404Scopus© Citations 5 - PublicationADAM10: a new player in breast cancer progression?(Cancer Research UK, 2015-08-18)
; ; ; ; Background: The ADAM proteases are best known for their role in shedding the extracellular domain of transmembrane proteins. Among the transmembrane proteins shed by ADAM10 are notch, HER2, E-cadherin, CD44, L1 and the EGFR ligands, EGF and betacellulin. As cleavage of several of these proteins has been implicated in cancer formation and progression, we hypothesised that ADAM10 is also involved in these processes. Methods: ADAM10 expression was decreased by RNA interference and the effects of this on cell numbers, invasion and migration were determined. We also examined the effect of ADAM10 inhibition on breast cancer cell line invasion and migration. Results: Using the triple-negative (TN) breast cancer cell lines, BT20, MDA-MB-231 and the non-TN cell line MDA-MB-453, knockdown of ADAM10 expression significantly decreased in vitro migration (Po0.01; for each cell line). Similarly, treatment with the ADAM10-selective inhibitor GI254023X reduced migration in the three cell lines (for BT20, Po0.001; for MDA-MB-231, P ¼ 0.005; for MDA-MB-453, P ¼ 0.023). In contrast, neither knockdown of ADAM10 nor treatment with the ADAM10-selective inhibitor GI254023X significantly affected cell numbers. Using extracts of primary breast cancers, higher levels of ADAM10 were found more frequently in high-grade vs low-grade tumours (Po0.001) and in oestrogen receptor (ER)-negative compared with ERpositive tumours (P ¼ 0.005). Analysis of pooled publicly available data sets found that high levels of ADAM10 mRNA were associated with adverse outcome in patients with the basal subtype of breast cancer. Conclusions: Based on our combined cell line and breast cancer extract data, we conclude that ADAM10 is likely to be involved in breast cancer progression, especially in the basal subtype.213Scopus© Citations 52 - PublicationAdvances in dynamic modeling of colorectal cancer signaling-network regions, a path toward targeted therapies(Impact Journals, 2014-12-31)
; ; ; ; ; ; The interconnected network of pathways downstream of the TGFβ, WNT and EGF-families of receptor ligands play an important role in colorectal cancer pathogenesis. We studied and implemented dynamic simulations of multiple downstream pathways and described the section of the signaling network considered as a Molecular Interaction Map (MIM). Our simulations used Ordinary Differential Equations (ODEs), which involved 447 reactants and their interactions. Starting from an initial "physiologic condition", the model can be adapted to simulate individual pathologic cancer conditions implementing alterations/mutations in relevant onco-proteins. We verified some salient model predictions using the mutated colorectal cancer lines HCT116 and HT29. We measured the amount of MYC and CCND1 mRNAs and AKT and ERK phosphorylated proteins, in response to individual or combination onco-protein inhibitor treatments. Experimental and simulation results were well correlated. Recent independently published results were also predicted by our model. Even in the presence of an approximate and incomplete signaling network information, a predictive dynamic modeling seems already possible. An important long term road seems to be open and can be pursued further, by incremental steps, toward even larger and better parameterized MIMs. Personalized treatment strategies with rational associations of signaling-proteins inhibitors, could become a realistic goal.234Scopus© Citations 15 - PublicationAdvances in mesenchymal stem cell-mediated gene therapy for cancerMesenchymal stem cells have a natural tropism for tumours and their metastases, and are also considered immunoprivileged. This remarkable combination of properties has formed the basis for many studies investigating their potential as tumour-specific delivery vehicles for suicide genes, oncolytic viruses and secreted therapeutic proteins. The aim of the present review is to discuss the range of approaches that have been used to exploit the tumour-homing capacity of mesenchymal stem cells for gene delivery, and to highlight advances required to realize the full potential of this promising approach.
548Scopus© Citations 84 - PublicationAICAR ameliorates high-fat diet-associated pathophysiology in mouse and ex vivo models, independent of adiponectinAims/hypothesis: In this study, we aimed to evaluate the therapeutic potential of 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), an activator of AMP-activated protein kinase, for ameliorating high-fat diet (HFD)-induced pathophysiology in mice. We also aimed to determine whether the beneficial effects of AICAR were dependent on adiponectin. Furthermore, human adipose tissue was used to examine the effect of AICAR ex vivo. Methods: Six-week-old male C57BL/6J wild-type and Adipoq −/− mice were fed a standard-fat diet (10% fat) or an HFD (60% fat) for 12 weeks and given vehicle or AICAR (500 μg/g) three times/week from weeks 4–12. Diet-induced pathophysiology was examined in mice after 11 weeks by IPGTT and after 12 weeks by flow cytometry and western blotting. Human adipose tissue biopsies from obese (BMI 35–50 kg/m2) individuals were incubated with vehicle or AICAR (1 mmol/l) for 6 h at 37°C, after which inflammation was characterised by ELISA (TNF-α) and flow cytometry. Results: AICAR attenuated adipose inflammation in mice fed an HFD, promoting an M1-to-M2 macrophage phenotype switch, while reducing infiltration of CD8+ T cells. AICAR treatment of mice fed an HFD partially restored glucose tolerance and attenuated hepatic steatosis and kidney disease, as evidenced by reduced albuminuria (p < 0.05), urinary H2O2 (p < 0.05) and renal superoxide levels (p < 0.01) in both wild-type and Adipoq −/− mice. AICAR-mediated protection occurred independently of adiponectin, as similar protection was observed in wild-type and Adipoq −/− mice. In addition, AICAR promoted an M1-to-M2 macrophage phenotype switch and reduced TNF-α production in tissue explants from obese human patients.Conclusions/interpretation: AICAR may promote metabolic health and protect against obesity-induced systemic diseases in an adiponectin-independent manner. Furthermore, AICAR reduced inflammation in human adipose tissue explants, suggesting by proof-of-principle that the drug may reduce obesity-induced complications in humans.
211Scopus© Citations 15 - PublicationAlternative Experimental Models of Ciliary Trafficking and Dysfunction in the Retina(Royal Society of Chemistry, 2019)
; ; ; The cilia of cells constituent to the retina are fundamental to vision. Of the ∼250 genes causative of inherited retinal degeneration, 20% mediate functions related to photoreceptor primary cilium formation, structure or function. Primary cilia are sensory and signalling organelles emanating from the plasma membrane of most cells. They regulate a variety of biological processes, including left/right body axis asymmetry, limb patterning, central nervous system development and sensation. Cilia function by forming a specialised region of plasma membrane which concentrates specific signalling components, such as for sonic hedgehog signalling and phototransduction. Here, we review the roles of ciliary signalling and trafficking pathways in retinal biology and disease with a focus on the potential of non-rodent, metazoan experimental models for shedding light on these processes.27 - PublicationAmyloid β-Protein Dimers Rapidly Form Stable Synaptotoxic Protofibrils(Society for Neuroscience, 2010-10)
; ; ; ; ; ; Nonfibrillar, water-soluble low-molecular weight assemblies of the amyloid beta-protein (A beta) are believed to play an important role in Alzheimer's disease (AD). Aqueous extracts of human brain contain A beta assemblies that migrate on SDS-polyacrylamide gels and elute from size exclusion as dimers (similar to 8 kDa) and can block long-term potentiation and impair memory consolidation in the rat. Such species are detected specifically and sensitively in extracts of Alzheimer brain suggesting that SDS-stable dimers may be the basic building blocks of AD-associated synaptotoxic assemblies. Consequently, understanding the structure and properties of A beta dimers is of great interest. In the absence of sufficient brain-derived dimer to facilitate biophysical analysis, we generated synthetic dimers designed to mimic the natural species. For this, A beta(1-40) containing cysteine in place of serine 26 was used to produce disulphide cross-linked dimer, (A beta S26C)(2). Such dimers had no detectable secondary structure, produced an analytical ultracentrifugation profile consistent for an similar to 8.6 kDa protein, and had no effect on hippocampal long-term potentiation (LTP). However, (A beta S26C)(2) aggregated more rapidly than either A beta S26C or wild-type monomers and formed parastable beta-sheet rich, thioflavin T-positive, protofibril-like assemblies. Whereas wildtype A beta aggregated to form typical amyloid fibrils, the protofibril-like structures formed by (A beta S26C)(2) persisted for prolonged periods and potently inhibited LIP in mouse hippocampus. These data support the idea that A beta dimers may stabilize the formation of fibril intermediates by a process distinct from that available to A beta monomer and that higher molecular weight prefibrillar assemblies are the proximate mediators of A beta toxicity.200Scopus© Citations 197 - PublicationAnalysis of Protein Phosphorylation Using Phos-Tag GelsPhos-tag gels are recent tools to dissect protein phosphorylation that operate by inducing a shift in the electrophoretic mobility of phosphorylated proteins compared to their nonphosphorylated counterparts. This article describes the preparation and electrophoresis of Zn2+ -Phos-tag gels along with electrotransfer of the separated phospho- and nonphosphoproteins onto a PVDF membrane using either wet-tank or semidry transfer. We also discuss the theory behind the technology with critical parameters to keep in mind for its successful application.
320Scopus© Citations 23 - PublicationAnalysis of Ras-effector interaction competition in large intestine and colorectal cancer context(Taylor & Francis, 2020-02-14)
; ; ; ; Cancer is the second leading cause of death globally, and colorectal cancer (CRC) is among the five most common cancers. The small GTPase KRAS is an oncogene that is mutated in ~30% of all CRCs. Pharmacological treatments of CRC are currently unsatisfactory, but much hope rests on network-centric approaches to drug development and cancer treatment. These approaches, however, require a better understanding of how networks downstream of Ras oncoproteins are connected in a particular tissue context–here colon and CRC. Previously we have shown that competition for binding to a ‘hub’ protein, such as Ras, can induce a rewiring of signal transduction networks. In this study, we analysed 56 established and predicted effectors that contain a structural domain with the potential ability to bind to Ras oncoproteins and their link to pathways coordinating intestinal homoeostasis and barrier function. Using protein concentrations in colon tissue and Ras-effector binding affinities, a computational network model was generated that predicted how effectors differentially and competitively bind to Ras in colon context. The model also predicted both qualitative and quantitative changes in Ras-effector complex formations with increased levels of active Ras–to simulate its upregulation in cancer–simply as an emergent property of competition for the same binding interface on the surface of Ras. We also considered how the number of Ras-effector complexes at the membrane can be increased by additional domains present in some effectors that are recruited to the membrane in response to specific conditions (inputs/stimuli/growth factors) in colon context and CRC.140Scopus© Citations 8 - PublicationThe analysis of serum response factor expression in bone and soft tissue prostate cancer metastases(Wiley, 2014-02)
; ; ; ; ; ; ; ; Castration-resistant prostate cancer (CRPC) represents a challenge to treat with no effective treatment options available. We recently identified serum response factor (SRF) as a key transcription factor in an in vitro model of castration resistance where we showed that SRF inhibition resulted in reduced cellular proliferation. We also demonstrated an association between SRF protein expression and CRPC in a cohort of castrate-resistant transurethral resections of the prostate (TURPS). The mechanisms regulating the growth of CRPC bone and visceral metastases have not been explored in depth due to the paucity of patient-related material available for analysis. In this study, we aim to evaluate SRF protein expression in prostate cancer (PCa) metastases, which has not previously been reported.395Scopus© Citations 15 - PublicationThe APC network regulates the removal of mutated cells from colonic crypts(Elsevier, 2014-04-10)
; ; ; ; ; Self-renewal is essential for multicellular organisms but carries the risk of somatic mutations that can lead to cancer, which is particularly critical for rapidly renewing tissues in a highly mutagenic environment such as the intestinal epithelium. Using computational modeling and in vivo experimentation, we have analyzed how adenomatous polyposis coli (APC) mutations and β-catenin aberrations affect the maintenance of mutant cells in colonic crypts. The increasing abundance of APC along the crypt axis forms a gradient of cellular adhesion that causes more proliferative cells to accelerate their movement toward the top of the crypt, where they are shed into the lumen. Thus, the normal crypt can efficiently eliminate β-catenin mutant cells, whereas APC mutations favor retention. Together, the molecular design of the APC/β-catenin signaling network integrates cell proliferation and migration dynamics to translate intracellular signal processing and protein gradients along the crypt into intercellular interactions and whole-crypt physiological or pathological behavior.240Scopus© Citations 16 - PublicationApplication of a Small EF Hand Affinity Tag for Expression, Purification and Biophysical Studies of G Protein-Coupled Membrane ReceptorsHeptahelical G protein-coupled receptors (GPCR) comprise a large family of integral membrane proteins involved in a wide array of cell signaling pathways. For high resolution structural studies of these receptors, multi-milligram quantities of pure and structurally unperturbed proteins are required. Purification of recombinant GPCRs typically involves their solubilization into detergent micelles followed by chromatographic purification. Because of relatively low expression levels of these recombinant receptors, it is challenging to design an efficient strategy for selective and efficient purification with high yield. Here, we describe a recently introduced purification system employing a high affinity molecular switch based on fragment complementation, with a calcium dependent capture and EDTA mediated chelation elution. This technique was successfully applied to the purification of the recombinant cannabinoid receptor CB2, a promising target for the development of drugs for inflammation, immunological disorders and pain. It is feasible that similar strategies can be successfully employed for expression and purification of other membrane protein targets.
281 - PublicationApplications of piezoresponse force microscopy in materials research: from inorganic ferroelectrics to biopiezoelectrics and beyondPiezoresponse force microscopy (PFM) probes the mechanical deformation of a sample in response to an electric field applied with the tip of an atomic force microscope. Originally developed more than two decades ago to study ferroelectric materials, this technique has since been used to probe electromechanical functionality in a wide range of piezoelectric materials including organic and biological systems. Piezoresponse force microscopy has also been demonstrated as a useful tool to detect mechanical strain originating from electrical phenomena in non-piezoelectric materials. Parallelling advances in analytical and numerical modelling, many technical improvements have been made in the last decade: switching spectroscopy PFM allows the polarisation switching properties of ferroelectrics to be resolved in real space with nanometric resolution, while dual ac resonance tracking and band excitation PFM have been used to improve the signal-to-noise ratio. In turn, these advances have led to increasingly large multidimensional data sets containing more complete information on the properties of the sample studied. In this review, PFM operation and calibration are described, and recent advances in the characterisation of electromechanical coupling using PFM are presented. The breadth of the systems covered highlights the versatility and wide applicability of PFM in fields as diverse as materials engineering and nanomedicine. In each of these fields, combining PFM with complementary techniques is key to develop future insight into the intrinsic properties of the materials as well as for device applications.
695Scopus© Citations 71 - PublicationAn Assessment of the Permeation Enhancer, 1-phenyl-piperazine (PPZ), on Paracellular Flux Across Rat Intestinal Mucosae in Ussing ChambersPurpose: 1-phenyl piperazine (PPZ) emerged from a Caco-2 monolayer screen as having high enhancement potential due to a capacity to increase permeation without significant toxicity. Our aim was to further explore the efficacy and toxicity of PPZ in rat ileal and colonic mucosae in order to assess its true translation potential. Methods: Intestinal mucosae were mounted in Ussing chambers and apparent permeability coefficient (Papp) values of [14C]-mannitol and FITC-dextran 4 kDa (FD-4) and transepithelial electrical resistance (TEER) values were obtained following apical addition of PPZ (0.6–60 mM). Exposed issues were assessed for toxicity by histopathology and lactate dehydrogenase (LDH) release. Mucosal recovery after exposure was also assessed using TEER readings. Results: PPZ reversibly increased the Papp of both agents across rat ileal and distal colonic mucosae in concentration–dependent fashion, accompanied by TEER reduction, with acceptable levels of tissue damage. The complex mechanism of tight junction opening was part mediated by myosin light chain kinase, stimulation of transepithelial electrogenic chloride secretion, and involved activation of 5-HT4 receptors. Conclusions: PPZ is an efficacious and benign intestinal permeation enhancer in tissue mucosae. However, its active pharmacology suggest that potential for further development in an oral formulation for poorly permeable molecules will be difficult.
520Scopus© Citations 15 - PublicationAtomic and Close-to-Atomic Scale Manufacturing: A Review on Atomic Layer Removal Methods Using Atomic Force MicroscopyManufacturing at the atomic scale is the next generation of the industrial revolution. Atomic and close-to-atomic scale manufacturing (ACSM) helps to achieve this. Atomic force microscopy (AFM) is a promising method for this purpose since an instrument to machine at this small scale has not yet been developed. As the need for increasing the number of electronic components inside an integrated circuit chip is emerging in the present-day scenario, methods should be adopted to reduce the size of connections inside the chip. This can be achieved using molecules. However, connecting molecules with the electrodes and then to the external world is challenging. Foundations must be laid to make this possible for the future. Atomic layer removal, down to one atom, can be employed for this purpose. Presently, theoretical works are being performed extensively to study the interactions happening at the molecule–electrode junction, and how electronic transport is affected by the functionality and robustness of the system. These theoretical studies can be verified experimentally only if nano electrodes are fabricated. Silicon is widely used in the semiconductor industry to fabricate electronic components. Likewise, carbon-based materials such as highly oriented pyrolytic graphite, gold, and silicon carbide find applications in the electronic device manufacturing sector. Hence, ACSM of these materials should be developed intensively. This paper presents a review on the state-of-the-art research performed on material removal at the atomic scale by electrochemical and mechanical methods of the mentioned materials using AFM and provides a roadmap to achieve effective mass production of these devices.
117Scopus© Citations 32 - PublicationATR inhibitors as a synthetic lethal therapy for tumours deficient in ARID1A(Springer Nature, 2016-12-13)
; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; Identifying genetic biomarkers of synthetic lethal drug sensitivity effects provides one approach to the development of targeted cancer therapies. Mutations in ARID1A represent one of the most common molecular alterations in human cancer, but therapeutic approaches that target these defects are not yet clinically available. We demonstrate that defects in ARID1A sensitize tumour cells to clinical inhibitors of the DNA damage checkpoint kinase, ATR, both in vitro and in vivo. Mechanistically, ARID1A deficiency results in topoisomerase 2A and cell cycle defects, which cause an increased reliance on ATR checkpoint activity. In ARID1A mutant tumour cells, inhibition of ATR triggers premature mitotic entry, genomic instability and apoptosis. The data presented here provide the pre-clinical and mechanistic rationale for assessing ARID1A defects as a biomarker of single-agent ATR inhibitor response and represents a novel synthetic lethal approach to targeting tumour cells.366Scopus© Citations 206 - PublicationBacterial Cholangitis in Autosomal Dominant Polycystic Kidney and Liver DiseaseTo describe first episodes of bacterial cholangitis complicating autosomal dominant polycystic kidney disease (ADPKD) and autosomal dominant polycystic liver disease (ADPLD) and to identify risk factors for cholangitis episodes among patients with ADPKD-associated polycystic liver disease (PLD). We searched the electronic medical records at our tertiary referral center for episodes of cholangitis in patients with ADPKD or ADPLD from January 1, 1996, through June 30, 2017. Cases were categorized as suspected or definite cholangitis by expert review. Clinical, laboratory, and radiologic data were manually abstracted. A nested case-control study was conducted to investigate risk factors for cholangitis in patients with ADPKD. We identified 29 cases of definite or suspected cholangitis complicating PLD (24 with ADPKD-associated PLD and 5 with ADPLD). Among patients with definite cholangitis in ADPKD-associated PLD (n=19) vs ADPLD (n=4), the mean ± SD age was 62.4±12.2 vs 55.1±8.6 years, and 9 (47.4%) vs 0 (0%), respectively, were male. The odds of gallstones (odds ratio [OR], 21.6; 95% CI, 3.17-927; P<.001), prior cholecystectomy (OR, 12.2; 95% CI, 1.59-552; P=.008), duodenal diverticulum (OR, 13.5; 95% CI, 2.44 to not estimable; P=.004), type 2 diabetes mellitus (OR, 6.41; 95% CI, 1.01 to not estimable; P=.05), prior endoscopic retrograde cholangiopancreatography (OR, 14.0; 95% CI, 1.80-631; P=.005), and prior kidney transplant (OR, 8.06; 95% CI, 1.72-76.0; P=.004) were higher in patients with ADPKD-associated PLD with definite cholangitis compared to controls. Gallstones, prior cholecystectomy, duodenal diverticulosis, type 2 diabetes mellitus, prior endoscopic retrograde cholangiopancreatography, and prior kidney transplant constituted risk factors for cholangitis among patients with ADPKD-associated PLD.
86 - PublicationBasic fibroblast growth factor modifies the hypoxic response of human bone marrow stromal cells by ERK-mediated enhancement of HIF-1α activity(Elsevier, 2014-05)
; ; ; ; ; Human bone marrow stromal cells (hBMSCs, also known as bone marrow-derived mesenchymal stem cells) are promising tools for the cellular therapy of human pathologies related to various forms of hypoxia. Although the current concepts of their clinical use include the expansion of hBMSC in standard cell culture conditions, the effect of the mitogen-driven ex vivo expansion on the adaptation to the hypoxic environment is unknown. Here, we provide data that the basic fibroblast growth factor (FGF2) enhances the induction of a wide range of hypoxia-related adaptive genes in hypoxic hBMSCs. We identified that the FGF2 signal is transmitted by the ERK pathway similar to that of hypoxia that also utilises the distal elements of the same signalling machinery including the extracellular signal-regulated kinase 1/2 (ERK1/2) and mitogen-activated protein kinase kinases (MEK1/2) in hBMSCs. We found that the simultaneous activation of ERK1/2 by FGF2 and hypoxia transforms the activation dynamics from oscillatory into sustained one. Activated ERKs co-localise with stabilised hypoxia inducible factor-1α (HIF-1α) followed by the reduction of its nuclear mobility as well as increased DNA binding capacity leading to the up-regulation of hypoxia-adaptive genes. Our findings indicate that the status of the ERK pathway has significant impacts on the molecular adaptation of hBMSCs to the hypoxic milieu.201Scopus© Citations 15