Figure 4: Antioxidant and Antiapoptotic Activities of TT-10 in CMs

(A) Intracellular reactive oxygen species (ROS) production was induced by methylglyoxal (100 μmol/l). n = 4 per group. (B) Western blot analysis of cleaved caspase 3. Starved CMs were treated with 10 μmol/l of each compound for 18 h. Next, 1 mmol/l methylglyoxal was added, and CMs were cultured for 6 h. n = 5 per group. (C) Western blot analysis of NRF2 and KEAP1. CMs were treated with 10 μmol/l of each compound for 6 h. n = 7. (D) Coimmunoprecipitation assay. The MYC3-NRF2 and HA2-KEAP1–overexpressing HEK293 cells were treated with TT-10 (10 μmol/l) for 2 h. Cell lysates were immunoprecipitated with mouse immunoglobulin G, anti-MYC-tag, or anti-HA-tag antibody. Cell lysates (input), IgG immunoprecipitates (IP), anti-MYC IP, and anti-HA IP were probed with anti-MYC-tag and anti-HA-tag antibodies. n.s. p > 0.05, **p < 0.01 versus the untreated control. #p < 0.05, ###p < 0.001 versus the methylglyoxal-treated control. Abbreviations as in Figure 1.