Fig. 4.

Inhibiting GRP75 decreases reactive oxygen species (ROS) and improves mitochondrial function.

A, Confocal microscopy detection of HL-1 cells stained with 2′,7′-dichlorofluorescein diacetate (DCFH-DA). Scale bar = 100 μm. B, Quantification of ROS by DCFH-DA intensity in (A). C, Confocal microscopy detection of HL-1 cells stained with Mito Sox Red. Scale bar = 100 μm. D, Quantification of mitochondrial ROS by Mito Sox Red in (C). E, Western blot detection of antioxidant stress protein manganese superoxide dismutase (Mn-SOD) levels in atrial myocytes. F, Quantification of protein levels in (E). G, JC-1 staining. Scale bar = 100 μm. H, Quantification of mitochondrial membrane potential (Δψm) of HL-1 cells by JC-1 aggregate–monomer ratio. Data represent the mean ± SEM (n = 4 independent experiments), ^#P < 0.01, ANOVA with Bonferroni post-test. I, Analysis of oxygen consumption rate (OCR). Oligomycin inhibits ATP synthase (J), FCCP uncouples oxygen consumption from ATP production (K), and AA + Retenone inhibits complexes I and III (L). Data represent the mean ± SEM (n = 6 independent experiments), *P < 0.05, ^#P < 0.01, ANOVA with Bonferroni post-test. . (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)