Contractile Activity Regulates Inducible Nitric Oxide Synthase Expression and
NOi Production in Cardiomyocytes via a FAK-Dependent Signaling Pathway
Figure 3
FAK regulates contraction-dependent activation of AKT. (a) NRVMs were treated with myosin II ATPase inhibitors (BDM or blebbistatin (Bleb)) or (b) L-type calcium channel blockers (nifedipine or verapamil) for 24 h. Cell extracts (50 μg total protein per lane) were then separated by SDS-PAGE and Western blotting with antibodies specific for FAK phosphorylated at Y577, total FAK, AKT phosphorylated at T308 and S473, and total AKT. (c) NRVMs were infected, (20 moi) with Adv-expressing shRNAs for luciferase (shLuc), FAK (shFAK), or PYK2 (shPYK2) and then maintained under control conditions for 72 h. Equal amounts of cell extracts (50 μg total protein per lane) were then separated by SDS-PAGE and Western blotting with antibodies specific for FAK phosphorylated at Y397, total FAK, AKT phosphorylated at T308 and S473, total AKT, total PYK2, and GAPDH.
