As described previously, H9C2 cardiomyocytes were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco Laboratories) supplemented with 10% fetal bovine serum (FBS) (Gibco Laboratories) and 1% penicillin/streptomycin in an atmosphere of 90% air and 10% CO2 at 37 °C (19 (link)). The experiment was performed with the control, H/R, low-dose (2.5 µM Art), medium-dose (5 µM Art), and high-dose (10 µM Art) groups. To explore the effect of the Art and FAK pathway inhibitor PF573228 on H/R injury, another experiment was performed with the control, H/R, H/R + PF573228 group, the H/R + PF573228 + Art (10 µM Art) group. Inhibitors that do not affect the morphology or viability of H9C2 cells were used. Hypoxia was established by culturing H9C2 cells for 4 hours at a constant temperature three-gas incubator with a mixture of 95% N2, 5% CO2, and 1% O2 at 37 °C. After that, the cells were reoxygenated in a normoxic chamber for 2 hours. A myocardial cell model (H9C2 cells) of hypoxia/reoxygenation (H/R) was established.
The FAK inhibitor PF573228 was obtained from Sigma-Aldrich (St. Louis, MO) and dissolved with dimethyl sulfoxide (DMSO) to a final concentration of <0.1% per assay.
The FAK inhibitor PF573228 was obtained from Sigma-Aldrich (St. Louis, MO) and dissolved with dimethyl sulfoxide (DMSO) to a final concentration of <0.1% per assay.
