Figure 5. Evaluation of the effects of the ACSL4/GLUT1 axis on cell proliferation, apoptosis and tumorigenesis in Huh-7 and SK-HEP-1 cells. (A, B) The mRNA and protein expression levels of GLUT1 were determined by RT-PCR and western blotting assays after cells were transfected with sh-GLUT1 or sh-NC, respectively (*P<0.05, **P<0.01, compared with the sh-NC group). Next, Huh-7 and SK-HEP-1 cells were transfected with OE-ACSL4 and/or sh-GLUT1 and subjected to the following assays. (C–E) Western blotting assays were used to assess the levels of O-GlcNAc, ACSL4 and GLUT1. (F, G) CCK-8 assay was carried out to test cell proliferation. (H) Flow cytometry assay was used to determine cell apoptosis. (I) An in vivo xenotransplantation assay was used to assess the effects of the ACSL4/GLUT1 axis on the tumour formation ability of Huh-7 and SK-HEP-1 cells. (C–I: *P<0.05, compared with control group; #P<0.05, compared with OE-ACSL4 group).