Fig. 2
From: Mitochondrial glutathione transporter SLC25A40 regulates macrophage cytokine production
SLC25A40 stabilizes Complexes I, II, and III in the mitochondria. BMDMs (0.5 × 106 cells/mL) were transfected with 50 nM negative control siRNA or Slc25a40 siRNA for 8 h on day 1. On day 2, the cells were left untreated (0) or treated with LPS (100 ng/mL) for 0, 1, 3, 4, 8, or 24 h. (A) Representative Western blot images of the OXPHOS proteins, SLC25A40, and β-actin (n = 3). (B-G) Protein levels of Complex I (B), II (C), III (D), IV (E), V (F), and SLC25A40 (G) over β-actin were measured by Western blotting and quantified by densitometry. (H, I) mRNA levels of Gclc (H) and Gclm (I) over housekeeping gene Rps18 were quantified by qPCR. (J, K) Cells were stained with CellROX (J) (n = 4) or MitoSOX (K) 30 min before harvesting and subjected to flow cytometry analysis. Data are means ± SEM. Results were obtained from at least three independent experiments. *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001 as calculated using two-way ANOVA for multiple comparisons or t-tests for comparisons between two groups.
