Fig. 8
Fluphenazine is an activator of KLF14 and inhibits HCC cells growth. A The chemical structure of Fluphenazine Hydrochloride. B The luciferase activity of reporters was analyzed in HepG2 cells transfected with pGL4-KLF14-luc constructs after 48 hours treatment with 10 μM Fluphenazine Hydrochloride or PBS. C HepG2 cells were transfected with pGL4-IRP2-luc plasmids for 12 hours and incubated with 10 μM Fluphenazine Hydrochloride or PBS for 48 hours. IRP2 promoter luciferase activity was then examined. D, E HepG2 and Huh7 cells were treated with PBS or Fluphenazine Hydrochloride at the indicated dosage for 48 h, mRNA (D) and protein levels (E) of KLF14 and IRP2 were examined by qRT-PCR and western blotting respectively. F Relative cellular LIP content of HepG2 and Huh7 cells were measured with the treatment of PBS or Fluphenazine Hydrochloride at the indicated dosage for 36 h. G The cell apoptosis rate of HepG2 and Huh7 cells with incubation of PBS or Fluphenazine Hydrochloride at the indicated dosage for 48 h were investigated by flow cytometry. H Cell growth curve of HepG2 and Huh7 cells were measured in media supplemented with PBS or Fluphenazine Hydrochloride at the indicated dosage for 4 days. All the results are presented as means ± SD from three independent experiments. Two-tailed unpaired Student’s T-tests were performed. n.s, not significant, *P < 0.05, **P < 0.01 and ***P < 0.001