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. 2021 Apr 19;11:8430. doi: 10.1038/s41598-021-87225-8

Figure 4.

Figure 4

Curcumin-mediated Nrf2 activation is dependent on the phosphorylation of p62 on S351. (A) HEK293 cells were transiently transfected with the Myc-Nrf2 expression plasmid, and then treated with DMSO (−) or 10 μM curcumin (+) for 12 h. The cell lysates were used for Nrf2 immunoprecipitation using an anti-Myc antibody. The protein level of Keap1 co-immunoprecipitated with Nrf2 was examined by immunoblotting using an anti-Keap1 antibody. Full blots are provided in Supplementary Fig. S11. (B) The bar graph shows the relative ratio of Keap1 against Nrf2 co-immunoprecipitated with the Myc antibody in cells treated with curcumin (CCM) or not. (C) HEK293 cells were transiently co-transfected with the ARE-Luc reporter and TK-Renilla plasmids along with the Myc-p62 wild-type or mutant (S349A) plasmid. After treatment with either DMSO (−) or 10 μM curcumin (+) for 12 h, the cells were assayed for the luciferase activity. Data shown are mean ± S.E. of three independent experiments and were analyzed using the Student’s t-test. (*p < 0.05, ***p < 0.001).