Fig. 3.

Nrf2 knockdown prevents NFT-dependent elevation of glutathione levels. a Nrf2, BiP (GRP78), or random targets (scrambled) were silenced in HepG2 cells by utilizing 40 μl of a 10 μM siRNA stock solution per 10⁶ cells for 24 h. Cells were then exposed to NFT (20 μM) for 48 h, and cell homogenates were analyzed by western blotting for the levels of Nrf2, GCLC, and GAPDH. For quantification, band intensities were normalized to those of GAPDH. Values of untreated control cells (w/o NFT) were defined as unity, and all other bands were expressed as fold change compared to control. b The siRNA-silenced cells were analyzed for their intracellular content of glutathione. Differences were tested for significance by one-way ANOVA, followed by Bonferroni’s post hoc test, *p < 0.05 for comparison of NFT treatment vs. untreated controls and for comparison of NFT treatment vs. siRNA knockdown. Data are means ± SD of three independent experiments. Individual values of the 3 biological replicates are indicated by dots