Figure 5
From: Pterisolic Acid B is a Nrf2 Activator by Targeting C171 within Keap1-BTB Domain
J19-1 actives Nrf2 pathway by inhibiting Keap1-mediated Nrf2 ubiquitination in vivo in a C171 dependent manner.
(a) HEK293 cells were transiently transfected with WT-Keap1 or C171S-Keap1, Gal4-Neh2 and HA-Ub and treated with indicated concentration of J19-1 or tBHQ for 5 h. Lysates were immunoprecipitated by anti-Gal4 antibody and ubiquitination was assessed using anti-HA antibody. (b) HEK293 cells were transiently transfected with HA-Cul3 and Flag-tagged WT (or C171S) Keap1 and then treated with tBHQ or 4 μM J19-1 for 5 h. Pull-down using Flag beads and ensuing immunoblot analysis with anti-HA antibody.
