Fig. 2 | Cell Death & Differentiation

Fig. 2

From: A p53/miR-30a/ZEB2 axis controls triple negative breast cancer aggressiveness

Fig. 2

p53 regulates miR-30a-5p and miR-30a-3p expression. a Ectopic p53 expression in MDA157 results in an augment of mature miR-30a-5p and miR-30a-3p levels. The immunoblot on the right shows p53 expression in MDA157 engineered cells. GAPDH was used as a loading control. b p53 silencing in HCT116 results in a decrement of both miR-30a-5p and miR-30a-3p. The extent of p53 silencing is shown in the right panel. c Ectopic p53 induces an increase of pri-miR-30a levels in MDA157. d Decrease of pri-miR-30a levels in p53-depleted HCT116 and MCF7 cells. ad Gray columns represent p53-modulated samples; black columns represent control cells. e p53 regulates the MIR30A promoter. Silencing of p53 results in a decrement of the MIR30A promoter activity (30wt). The mutagenesis of the two p53 binding sites, singularly (30mut1, 30mut2) or in combination (30mut1/2), abrogates this effect. Results represent the mean value of three independent experiments ± SD. f p53 binds the miR-30a promoter. Chromatin immunoprecipitation was performed with the DO-1 anti-p53 monoclonal antibody on HCT116 genomic DNA. Isotype-matched pre-immune mouse IgG was used as a negative control. The immunoprecipitated chromatin was assayed for the enrichment of the target MIR30A promoter (miR30-1 and 2, the regions encompassing the two p53BS) by qPCR. The p53 binding region of the p21 promoter and an irrelevant genomic region (CTR neg) [59] were used as positive and negative control, respectively. Data are reported as fold enrichment over control samples (immunoprecipitation with pre-immune IgG) p values were calculated by two-tailed t-test; *p < 0.05, **p < 0.01.

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