Fig. 5

p62 stabilizes Nrf2 in hFOB 1.19 cells exposed to CNR2 agonists. hFOB 1.19 cells were cultured at 34 °C until reaching confluence and then transfected with empty vector or p62 overexpression vector (SQSTM1 OE). Then, culture medium was replaced, and cells were then cultured at 39 °C in presence of 50 nM HU308, 10 μM JWH133 for 48 h. a-b The protein levels of p62 and Keap1 were analyzed with western blotting. c Post a 48-h transfection, hFOB 1.19 cells were harvested at 0.5, 1, 3, 6, 8 h following treatment of 100 μg/mL cycloheximide. The expression levels of Nrf2 were determined with western blotting analysis. d-e For Co-IP assay, beads coated with anti-Keap1 was used to pull down intracellular Keap1, and the expression levels of Keap1 and Nrf2 were determined with western blotting