Fig. 5.

IRP2 is the target gene of KLF14 to regulate cellular iron content. A HepG₂ cells with steady expression of KLF14-WT-3 × Flag or vector (control) were grafted into nude mice subcutaneously. The effects of KLF14 overexpression and/or iron supplementation on tumor growth in vivo was then assessed. n = 6. B Tumors were isolated and weight was measured. n = 6. C Relative mRNA and protein levels of KLF14, IRP2, TfR1, FH and GAPDH (as control) were examined by qRT-PCR and western blotting respectively. n = 6. D Immunohistochemistry staining for H&E, Perl’s Blue (iron), Ki67, cleaved caspase3 and IRP2 in tumor tissues isolated from mice. Scale bar, 50 μm. E Relative mRNA level of IRP2 in 69 paired tumor tissues and adjacent tissues of HCC. F The correlation analysis of KLF14 and IRP2 expression was conducted based on the mRNA levels of tumor tissues in HCC (n = 69). G Statistics of IHC staining displayed the percentages of HCC specimens with higher or lower KLF14 expression and corresponding IRP2 levels. H Representative images of KLF14 staining and corresponding IRP2/TfR1 staining were shown (scale bar, 100 μm). I Immunohistochemistry staining of KLF14, IRP2 and Perl’s Blue (iron) in HCC tumor and adjacent tissues (n = 4) were exhibited (scale bar, 50 μm). Two-tailed unpaired Student’s T-tests were performed. n.s, not significant, *P < 0.05, **P < 0.01 and ***P < 0.001