Fig. 3
Vitexin regulated the expression of Keap1/NRF2/HO-1 in vivo and in vitro. (A) Predicted binding mode of vitexin with Keap1. (B) Viability of HK2 cells treated with vitexin. (C) Total NRF2 protein levels in HK2 cells pretreated with or without vitexin, as detected by WB. The ubiquitination of NRF2 was examined by immunoprecipitation assays. (D) Representative fluorescence images showing the colocalization of NRF2 and nuclei in HK2 cells incubated with vitexin (100 µmol/L). (E) Quantification of HO-1 mRNA expression in vitexin-treated HK2 cells. (F) Schematic IHC representation of NRF2 in vehicle- and vitexin-treated mouse kidney tissues and quantified images are shown. (G) Quantification of HO-1 mRNA expression in vitexin-treated mice kidneys. (H) NRF2 protein expression was measured by western blot. Lamin B was used as an internal control for nuclear proteins, and β-actin was used as an internal control for cytoplasmic proteins. The data are shown as the mean ± S.D. (n = 3). ****P < 0·0001, **P < 0·01, *P < 0·05 (one-way ANOVA for B & G, t-text for E & F)