FIG 4.

Establishment of sMaf-deficient MEFs expressing the T-N2G protein. (A) Schematic diagram of the protocol for obtaining stable cell lines. After transfection followed by puromycin selection, mock and T-N2G MEFs were established. (B) Mock, T-N2G, and F0G2K0-T MEFs were treated with 100 μM DEM for 5 h, and their nuclear lysates were analyzed by immunoblot analysis using an anti-Nrf2 antibody. Lamin B1 was detected as a loading control. (C) T-N2G-mediated induction of typical Nrf2 target genes, Nqo1, Gsta4, and Gclc, was examined by qPCR. The expression level of each mRNA was normalized to that of Hprt mRNA. The data represent the means ± SDs (n = 3). The gene expression levels in vehicle-treated mock cells were set to 1.