Fig. 3

miR-29a expression is induced by P53. A A schematic representation showed a potential P53 binding site 2.2 kb upstream of the miR-29a TSS. B A sequence of the miR-29a promoter with the P53 binding site or site directed mutagenesis of the identified sequence was cloned into a pEZX-MT01 luciferase reporter vector, after transfection into glioma cells (U87 and U251). The direct regulation of P53 on miR-29a was assessed by luciferase reporter assay; the pEZX basic vector was used as a control. C After p53 was effectively knocked down in glioma cells (U87 and U251), miR-29a expression was detected by relative quantity RT-PCR. D The wild-type pEZX-miR-29a construct-transfected cells with or without p53 knockdown was treated and then the luciferase reporter activity in these cells was detected. E Commercial soluble p53 protein was added to culture media and miR-29a expression in glioma cells was measured. Results show the mean ± SD (n = 3). *P < 0.05; **P < 0.01