Fig. 4

Resisting C-2 induced apoptosis by p62 activated Nrf2 pathway in early time. a Western blotting assay was used to detect the expression levels of p62, Nrf2, Keap1 and NQO1 after treated with 4 μM of C-2 for 24 h in BIU87 and EJ cells. b BIU87 cells were treated with C-2 for 6 h. The treated and untreated samples were stained with Nrf2 antibody (Green) and p62 antibody (Red) and DAPI (Blue) (magnification, 400X). The arrow was indicating Nrf2 nuclear translocation. c Immunoprecipitation assay showed the effect of C-2 on the binding of p62 and Keap1 proteins in BIU87 cells for 6 h and 12 h. d Western blotting assay showed the effect of p62 siRNA (20 nM) on expression of p62 and p-Nrf2 proteins in BIU87 cells. e MTT assay detected the effect of siRNA targeting to p62 on the survival rate of BIU87 cells incubated with 4 μM of C-2 for 6 h, **P < 0.02 vs C-2 treatment group