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. 2017 Feb 28;50(2):91–96. doi: 10.5483/BMBRep.2017.50.2.188

Fig. 3.

Fig. 3

p62 is required for verapamil-induced Keap1 degradation and Nrf2 activation. (A) p62+/+ or p62−/− MEF cells were incubated in the absence or presence of verapamil (100 μM) for 18 h. Lysates of p62+/+ or p62−/− MEF cells were subjected to immunoblot analysis with antibodies against Keap1, p62, and β-actin (loading control). (B) Densitometric analysis of Keap1 immunoblots was obtained in (A). Total RNA isolated from cells treated as described in (A) was subjected to qRT-PCR analysis for mRNAs of Keap1 (C), GSTA1 (D), HO-1 (E), and Nqo-1 (F). (G) Hepa1c1c7 cells transfected with control siRNA (si-control) or p62 siRNA were treated with DMSO or verapamil (100 μM) for 18 h. The cells were lysed and subjected to immunoblot analysis with antibodies against Keap1, p62, and β-actin (loading control). (H) Densitometric analysis of Keap1 immunoblots was obtained in (G). qRT-PCR analysis of mRNAs of Keap1 (I), GSTA1 (J), HO-1 (K), and Nqo-1 (L). Data are presented as the means ± SD from three independent experiments. *P < 0.05.