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. 2019 Jul 16;42(3):1005–1016. doi: 10.3892/or.2019.7235

Figure 3.

Figure 3.

Gene expression and DNA CpG methylation analysis of selected genes in SV-HUC-1 cells. (A and B) Gene expression analysis. After 0.5 µM sodium arsenite treatment for 14 days, RNA was extracted and analyzed by (A) RT-PCR, or protein was extracted and analyzed by (B) western blotting with anti-WIF1 antibodies. The quantified data of the histograms were calculated from seven (WIF1 mRNA) and four (WIF1 protein) independent experiments. ***P<0.001 compared to control. (C) DNA CpG methylation analysis of the WIF1 gene. After 0.5 µM sodium arsenite treatment for 14 days, DNA CpG methylation was analyzed by BSP-MSP/USP assay. The MSP ratio=MSP intensity/MSP intensity + USP intensity. (D and E) Gene expression and DNA CpG methylation analysis of the WIF1 gene after 0.5 µM sodium arsenite treatment for 10 weeks. (D) The mRNA expression level of WIF1 was analyzed by RT-PCR (n=3 from 3 independent experiments; ***P<0.001 compared to control) and (E) DNA CpG methylation was analyzed by BSP-MSP/USP assay. WIF1, Wnt inhibitory factor 1; BSP, bisulfite-sequencing PCR; MSP, methylation-specific PCR; USP, unmethylation-specific PCR.