Fig. 2. H2S-induced mitochondrial uncoupling requires SQR and is separated from the inhibition of complex IV by H2S.
(A and B) Expression levels of SQR protein (A) and mRNA (B) in mouse primary microglia (Mic), neurons (Neu), and astrocytes (Ast). Liver: positive control. (C to G) ADT-OH failed to induce mitochondrial uncoupling in primary neurons, as indicated by real-time change and statistical analysis of OCR (C and D; n = 6), ATP levels (E; n = 3), and ΔΨm (F, representative images of JC-1 staining; G, statistical analysis; n = 9). Ctrl, Veh-treated neurons. (H to J) Real-time change and statistical analysis of OCR (H and I; n = 9), and ATP levels (J; n = 3) in primary neurons infected with lentivirus expressing SQR (LV-SQR) or control lentivirus (LV-Ctrl) and then treated with ADT-OH. (K to M) Real-time change and statistical analysis of OCR (K and L; n = 7), and ATP levels (M; n = 3) in primary microglia infected with lentivirus expressing SQR-shRNA (short hairpin RNA) or nonsense shRNA (Ctrl-shRNA) and then treated with ADT-OH. Ctrl, cells without lentiviral infection. (N) Complex IV activity in BV2 microglia treated with ADT-OH. (O) Complex IV activity in WT HEK, Ctrl HEK, and CBS HEK cells; n = 3.