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Fig. 7 | BMC Medicine

Fig. 7

From: Nitazoxanide inhibits acetylated KLF5-induced bone metastasis by modulating KLF5 function in prostate cancer

Fig. 7

Nitazoxanide attenuates the upregulation of MYBL2 expression by Ac-KLF5 in prostate cancer cells. a Expression of MYBL2 and TIMM8A in PC-3 and DU 145 cells expressing different forms of KLF5, as determined using qPCR. C4-2B and parental PC-3 cell lines were used as controls, and GAPDH was used as a loading control. Statistical analysis was performed using one-way ANOVA with multiple comparisons. b Cells were transfected with 50 nM siKLF5 concomitantly with or without NTZ (5 μM) treatment for 24 h and then processed real-time qPCR. Statistical analysis was performed using one-way ANOVA with multiple comparisons. c NTZ downregulated MYBL2 expression in PC-3-KQ cells and DU 145-KQ cells at mRNA levels, as detected by qPCR. NTZ treatments were at indicated concentrations for 48 h. Statistical analysis was performed using one-way ANOVA with multiple comparisons. d NTZ treatment in mice reduced MYBL2 expression in PC-3-KQ bone metastasis, as detected by IHC staining and indicated by IHC images and intensities of staining signals. n = 5 tumors for each group. Student’s t-test was performed. Scale bar, 200 μm. e The MYBL2 gene promoter contained multiple potential KLF5 binding sites, as predicted by the JASPAR program. f, g NTZ had different effects on the binding of Ac-KLF5 to different Ac-KLF5-binding sites of the MYBL2 gene promoter, as determined by ChIP and regular RT-PCR (f) or real-time qPCR (g) using PC-3-KQ cells treated with NTZ for 48 h and PC-3-KR cells. All bar graphs are shown as mean ± SD of three independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ns, no statistically significant

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