Fig. 4

Ectopic expression and knockdown of KRAL. a qRT-PCR data showing the relative expression of KRAL in HepG2/5-FU and SMMC-7721/5-FU cells 48 h after transfection with lentivirus expressing empty vector or KRAL, *p < 0.05. The mRNA and protein levels of Keap1 in control and KRAL-overexpressing cells were determined by RT-PCR and western blotting, respectively. GAPDH was used as a reference; data are expressed as the mean ± SD; bar graph indicates the normalized values from at least 3 separate experiments; *p < 0.05 vs control group. b qRT-PCR data showing the relative expression of KRAL in HepG2 and SMMC-7721 cells 48 h after transfection with lentivirus expressing shRNA against scramble or KRAL, *p < 0.05, ^#p < 0.05. The mRNA and protein levels of Keap1 in control and KRAL knockdown cells were determined by RT-PCR (*p < 0.05, ^#p < 0.05) and western blotting (*p < 0.05, ^#p < 0.05), respectively. GAPDH was used as a reference; data are expressed as the mean ± SD; bar graph indicates the normalized values from at least 3 separate experiments. c Cellular fractions were isolated from HepG2 and SMMC-7721 cells. KRAL was mainly distributed in the cytoplasm. GAPDH mRNA and U6 RNA were used as controls for the cytoplasmic and nuclear RNA fractions, respectively. Data are expressed as the mean ± SD; columns: mean of three independent experiments