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. 2018 Dec 20;9(1):26–34. doi: 10.1002/2211-5463.12554

Figure 2.

Figure 2

p53‐mediated otop2 transcriptional regulation. The mRNA and protein expression of p53 and OTOP2 in CRC cell lines expressing either wtp53 or mtp53 using real‐time PCR (A), WB (B) and IF assay with 200× amplification (C). Three putative p53 consensus binding sequences of the otop2 promoter (CBS1‐3) were inserted upstream of the minimal SV40 promoter in the pGL‐3 luciferase reporter plasmid cotransfected with wtp53 or mtp53 (R273H and P309S), respectively, in SW403 (p53 null) cells. The p53‐dependent activity on CBS at the otop2 promoter region demonstrated using an imaging reporter assay (D). The binding status between endogenous p53 and the CBS1‐3 of the otop2 promoter in wtp53 and mtp53 CRC cells using ChIP assay (E). All the data are presented as mean ± SEM of triplicate individual experiments. “*” represents p value less than 0.05 by one‐way ANOVA.