Knockdown of UQCRC2 and exposure to polyunsaturated fatty acid prevents apoptosis in cNLRX1 cells. (A) Western blot analysis of UQCRC2 expression in cEmpty and cNLRX1 cells treated with siGFP or siUQCRC2, as indicated. Relative expression (Rel. expr.) for UQCRC2 was determined by densitometric analysis for UQCRC2 and respective β actin bands. Protein molecular weight is indicated in kilodaltons. (B and C) Cells were treated with the indicated siRNA and subjected to hypoxia followed by reperfusion for 24 h. (B) Cleaved caspase-3 staining and (C) MitoSOX Red were analyzed by digital image analysis for normoxia (white bars) and reoxygenation (black bars) samples. (D) Cleaved caspase-3 staining was analyzed by digital image analysis for normoxia (white bars) and reoxygenation (black bars) during 24 h of exposure to the indicated fatty acids. Data are expressed as mean ± SEM and were analyzed using an unpaired t test (*, P < 0.05; **, P < 0.01; ***, P < 0.001 vs. normoxic control). All conditions were determined in triplicate; representative data from three independent experiments are shown.