Fig. 8.

KRAL reverses 5-FU resistance in HCC cell lines by regulating Keap1 expression. a Western blotting was performed to detect Keap1 expression in KRAL-overexpressing HepG2/5-FU and SMMC-7721/5-FU cells transfected with Keap1 siRNA or miR-141 mimics. 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, ^#p < 0.05, ^&p < 0.05. b The IC₅₀ of 5-FU in KRAL-overexpressing HepG2/5-FU and SMMC-7721/5-FU cells transfected with Keap1 siRNA or miR-141 mimics. Data are expressed as the mean ± SD; columns: mean of three independent experiments; *p < 0.05, **p < 0.01. c Transfected HepG2/5-FU and SMMC-7721/5-FU cells were exposed to the indicated doses of 5-FU for 48 h. The cells were stained with annexin-V/PI and subjected to flow cytometry. Bar graph indicates the relative percentages of apoptotic cells from three independent experiments, *p < 0.05. d The IC₅₀ of 5-FU in KRAL-silenced HepG2 and SMMC-7721 cells transfected with Keap1-FLAG plasmids or miR-141 inhibitor. Data are expressed as the mean ± SD; columns: mean of three independent experiments; *p < 0.05, **p < 0.01. e KRAL-silenced HepG2 and SMMC-7721 cells were exposed to the indicated doses of 5-FU for 48 h. The cells were stained with annexin-V/PI and subjected to flow cytometry. Bar graph indicates the relative percentages of apoptotic cells from three independent experiments. *p < 0.05, **p < 0.01