Fig. 7.

Mutation of KLF14 results in minimal change of HCC cells growth. A, B ChIP assay revealed downregulation effects of KLF14 on H3K9ac (A) and H4K16ac (B) of IRP2 promoter. KLF14-WT-3 × Flag and KLF14-M4–3 × Flag overexpressed HepG₂ cells were harvested, the samples were immunoprecipitated with anti-H3K9ac and anti-H4K16ac antibody and the precipitated DNA fragments were subjected to qRT-PCR in the IRP2 promoter regions. C, D Relative mRNA (C) and protein levels (D) of IRP2, TfR1, FH and GAPDH (control) in KLF14-M4–3 × Flag overexpressed HepG₂ cells. E, F Relative cellular LIP content in KLF14-M4–3 × Flag overexpressed cell lines were measured at 24 h (E) and 36 h (F). G The cell apoptosis rate of KLF14-M4–3 × Flag overexpressed cells were investigated by flow cytometry. H, I Cell growth curve (H) and Colony formation ability (I) of KLF14-M4–3 × Flag overexpressed cells. J Schematic diagram depicted the working model of KLF14. It shows that KLF14 inhibits the transcription of IRP2 via recruiting SIRT1 which then causes TfR1 downregulation and ferritin upregulation on the basis of IRP-IREs system. This results in cellular iron deficiency and suppression of HCC cells growth. 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