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Fig. 9 | Cell Communication and Signaling

Fig. 9

From: lncRNA KRAL reverses 5-fluorouracil resistance in hepatocellular carcinoma cells by acting as a ceRNA against miR-141

Fig. 9

KRAL inhibits the Nrf2 pathway by regulating Keap1 expression in a dose-dependent manner. a Western blotting was performed to detect the protein levels of Nrf2 and HO-1 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 vs the CON group, #p < 0.05 vs the KRAL group, &p < 0.05 vs the KRAL+siKEAP1 group. b Western blotting was performed to detect the protein levels of Nrf2 and HO-1 in KRAL-silenced HepG2 and SMMC-7721 cells transfected with Keap1-FLAG plasmids or miR-141 inhibitor. 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 the shcon group, #p < 0.05 vs the sh2 group, &p < 0.05 vs the sh2 + Keap1 group. c HepG2/5-FU and SMMC-7721/5-FU cells were transiently transfected with HO-1-ARE-luciferase plasmid or control vector and then transfected with different concentrations of KRAL or KRAL-mut plasmids in the presence or absence of miR-141 mimics; tBHQ was used as a positive control. The induced fold change in luciferase activity for cell lysates was analysed by normalizing the transfection efficiency and dividing the values of each experiment to those of the control. Data are expressed as the mean ± SD; columns: normalized mean of three independent experiments; *p < 0.05 vs the 0 nm group, #p < 0.05 vs the 20 nm group, &p < 0.05 vs the 40 nm group, Δp < 0.05 vs the 80 nm group. d HepG2 and SMMC-7721 cells were transiently transfected with HO-1-ARE-luciferase plasmid or control vector and then transfected with different concentrations of shRNA-KRAL or KRAL-mut plasmids in the presence or absence of miR-141 inhibitors; tBHQ was used as a positive control. The induced fold change in luciferase activity for cell lysates was analysed by normalizing the transfection efficiency and dividing the values of each experiment relative to those of the control. Data are expressed as the mean ± SD; columns: mean of three independent experiments; *p < 0.05 vs the 0 nm group, #p < 0.05 vs the 20 nm group, &p < 0.05 vs the 40 nm group, Δp < 0.05 vs the 80 nm group

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