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. 2019 May 23;11:1758835919850665. doi: 10.1177/1758835919850665

Figure 5.

Figure 5.

p53 directly induced miR-193a and epidermal growth factor receptor (EGFR) to form a double-negative feedback loop. (a) Schematic illustrating the putative p53/EGFR-binding sites in the miR-193a promoter. (b) Direct binding of p53 and EGFR to promoter regions of miR-193a as indicated by PCR-based chromatin immunoprecipitation (ChIP) assays. Binding was confirmed by semi-quantitative PCR, followed by gel electrophoresis. (c) Luciferase reporter assays confirmed the suppression of the miR-193a promoter by EGFR, and the promotion of the miR-193a promoter by p53 through the potential binding. (d) qRT-PCR analysis of the miR-193a levels in A549 cells after the knockdown or overexpression of p53 or EGFR. (e) Quantitative RT-PCR showing the miR-193a levels in non-small cell lung carcinoma (NSCLC) cell lines, i.e., A549: p53 WT; H1299: p53 Null. (f, g) Western blot analysis of p53 and EGFR protein levels in A549 and H1299 cells. (f) Representative images, (g) quantitative analysis. (h) Western blot analysis of p53 and EGFR levels in A549 cells transfected with control siRNA, p53 siRNA, p53 siRNA plus miR-193a mimic, control vector, p53 vector, or p53 vector plus miR-193a inhibitor. *p < 0.05; **p < 0.01; ***p < 0.001.