Mouse monoclonal antibody against gapdh

hMSCs were encapsulated and cultured in adhesive peptide-functionalized hydrogels overnight. Cells were washed twice with PBS and lysed by sonication on ice in cell extraction buffer containing protease and phosphatase inhibitors (ThermoFisher). The lysates were cleared by centrifugation at 10,000 rpm for 15 min at 4 °C and the extract was stored at −80 °C until analysis. Protein concentration was determined using a micro BCA kit (Pierce). Equivalent amounts of reduced, boiled (10 min at 70 °C) lysate were loaded on Bolt 10% Bis-Tris Plus gels (ThermoFisher) and subsequently transferred onto PVDF membranes. Membranes were probed with mouse monoclonal antibody against GAPDH (Abcam), mouse monoclonal antibody against FAK and rabbit polyclonal antibody against FAK [pY397] (ThermoFisher) at a 1:1000 dilution in 5% BSA TBS-T solution followed by fluorescent secondary antibodies (Li-Cor). Immunoblots were visualized on a Li-Cor Odyssey imaging system and analyzed using Image Studio Lite (Li-Cor) (Supplementary Fig. 20).

VEGF-A was acquired from Peprotech (Rocky Hill, NJ, USA); PGE2 came from Cayman Chemical (Ann Arbor, MI, USA); mouse monoclonal antibodies against cPLA₂ were purchased from Santa Cruz (Santa Cruz, CA, USA); mouse monoclonal antibody against GAPDH was purchased from Abcam (Cambridge, UK); rabbit polyclonal antibodies against phospho-cPLA₂ and COX-2 were purchased from Cell Signaling Technology (Danvers, MA, USA); secondary goat anti-mouse TEXAS RED conjugated antibody was purchased from Calbiochem; and secondary goat anti-rabbit IRDye 680 conjugated antibody was purchased from LI-COR (Lincoln, Dearborn, MI, USA). Cisplatin and dexamethasone were purchased from Sigma-Aldrich (Munich, Germany). Media, antibiotics and other reagents for cell cultures came from Innoprot (Elexalde Derio, Spain).