Figure 7.
The effects of H2S on Keap-1 ubiquitylation might contribute to promoting disulphide formation of Keap-1. H9C2 cells were treated with HG+Pal, HG+Pal+NaHS and HG+Pal+NaHS+DTT (20 μM, an inhibitor of disulphide bonds) for 48 h. (a) The ubiquitylation level of Keap-1 in mouse myocardia was detected by immunoprecipitation and western blotting. (b) The expression of Keap-1 was detected by western blotting. (c) The autophagosome was detected using an MDC test in H9C2 cells (green); scale bar: 100 μm. (d) The ratio of LC3 II to LC3 I was detected by western blotting. (e) The interaction of Keap-1 with P62 and LC3 II was detected by immunoprecipitation and western blotting. Values are presented as the mean±S.D. from n=3 replicates. *P<0.05 compared with the Ctl group; ##P<0.01 compared with the HG+Pal group; &&P<0.01 compared with the NaHS+DTT group