Fig. 3

Analysis of the promoter of MT1G and its induction by sorafenib. a Luciferase activity from different constructions derived from the promoter of MT1G. The constructions -1000, -416, -133, -28 nt and control (empty vector) were transfected into Huh7 cells, exposed to sorafenib 10 μM for 18 h. *: p < 0.05 compared to control. n.s.: non-significant compared to control. b analysis of the effect of NAC on the transcriptional activation of the 1-133 region of MT1G promoter by sorafenib. Luciferase activity was evaluated from Huh7 cells transfected with the 1-133 construction and the empty vector. Cells were preincubated with 10 mM NAC shortly before transfection and luciferase activity was measured after 18 h. The values of luciferase activity are normalized to the conditions without NAC, and expressed as a fold-induction with NAC. *: p < 0.05 compared to control. c Western blot with NRF2, ERK, pERK and Actin antibodies on Huh7 cells treated for 18 h with sorafenib 10 μM and/or NAC 10 mM (preincubated 1 h). d MT1G mRNA levels were analysed in Huh7 cells transfected with control siRNA vs siRNA directed against NRF2, and exposed to sorafenib 10 μM for 18 h as indicated. *: p < 0.05 compared to control siRNA treated with sorafenib