Focal adhesion complexes (FACs) are large multi-protein structures anchored at the plasma membrane of cells to connect the extracellular matrix to the cytoskeleton. Due to this membrane-spanning location, FACs have predominantly been associated with mechanosensing signalling pathways, particularly in skeletal muscle where FAC-related proteins are loading-responsive(1). Recently, however, FACs have also been implicated in the regulation of mechanistic target of rapamycin complex 1 (mTORC1) activity, a primary regulator of cell anabolism, in response to amino acids (AAs) and/or growth factors (GFs)(2). Importantly, however, these investigations were conducted in non-muscle cell types and therefore FACs role in AA and/or GF-mediated mTORC1 activation in skeletal muscle is yet to be elucidated. Our recent data displayed that mTORC1 activation occurs in close proximity to FACs following anabolic stimuli in human skeletal muscle(3) suggesting signalling responses to elevated AA/GF availability may centre at these complexes in vivo. While this displayed the localisation of mTORC1 activity at FACs, further studies specifically assessing the importance of FACs in the regulation of anabolic responses to AAs and/or GFs are required. Therefore, in this investigation we utilised two pharmacological compounds known to disrupt FACs (Y-27632 dihydrochloride – Rho-associated protein kinase inhibitor (50µM), Cilengitide – integrin antagonist (2.5µM)) and studied their effects on anabolic signalling responses to elevated AA (2x minimal essential medium (MEM) concentrations) or GF (10ng/mL IGF-1) concentrations. Immortalised human primary myotubes (C25 cell line) were proliferated and differentiated before being starved of nutrients (EBSS) for 4h in the presence of each compound, followed by a 1h period of AA/GF stimulation. Myotubes were then collected, lysed and prepared for immunoblotting (n=8 for each condition) for a variety of mTORC1-related signalling targets. Independent t-tests were used to test for statistical significance between each compound and an untreated (control) condition (significance set at p<0.05). In response to elevated AAs, Y-27632 reduced RPS6^Ser240/244 and RPS6^Ser235/236 phosphorylation compared to control (~36% & ~67% respectively, p<0.01) whilst cilengitide reduced RPS6^Ser235/236 phosphorylation (~34%, p=0.015) and elevated eEF2^Thr56 phosphorylation (~50%, p=0.012), all indicative of impaired mTORC1 activity. In response to IGF-1, phosphorylation at both sites on RPS6 was reduced by Y-27632 (RPS6^Ser240/244 – ~39%, RPS6^Ser235/236 – 68%, p<0.01), whereas cilengitide had no effect. Further markers of mTORC1 activity (p-4EBP1^Thr37/46), GF signalling (p-AKT^Ser473) and autophagy (LC3b II/I ratio) were unaltered by the FAC inhibitors in both conditions. Importantly, abundance of a prominent FAC protein (Talin1) and mTOR itself were also unaltered suggesting the effects of Y-27632 and cilengitide occurred independently of changes to FAC or mTOR content. These results show that intact FACs are required for some, but not all, mTORC1-related signalling responses to elevated AAs and GFs in human skeletal muscle cells, which builds on our current understanding of anabolic regulation in this tissue. Further work will employ immunofluorescent staining to confirm FAC disruption and the SUnSET technique to determine effects of global protein synthesis.