Fig. 4.

SD2267 promoted the nuclear translocation of NRF2 and activated NRF2 target genes in AML12 cells

(A) AML12 cells were incubated with 300 nM of SD2267 for the indicated times, and subjected to nuclear fractionation. GAPDH was used as a loading control for cytosolic proteins, and Lamin B was used as a loading control for nuclear proteins. Protein expression levels were determined by western blotting, and densitometry was performed using three independent experiments. **p < 0.01, ***p < 0.001. (B) AML12 cells were incubated with 100 or 300 nM of SD2267 for 3 h, and subjected to NRF2 immunofluorescence staining (green). DAPI was used to stain nuclei (blue). In merged NRF2 and DAPI images, nuclear NRF2 stained blue-green (Scale bar = 25 μm). (C) AML12 cells were incubated with 100 nM of SD2267 for the indicated times. Gene expression levels of HMOX1, NQO1, GCLC, and GCLM were measured by qRT-PCR. Relative expression levels are presented as mean ± SEM (n = 3 per group). **p < 0.01, ***p < 0.001, ns; not significant versus the control. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)