miR-24-3p affects the p53 pathway by regulating the expression of PRKCH.

(A) Western blot assays tested the effect of PRKCH on the expression of p53 and p21 in ACC-M cells (left). Western blot assays determined that the overexpression of p53 influenced the p21 protein in ACC-M cells. (B) The effect of miR-24-3p on the expression of p53 and p21 in ACC-M cells was detected by western blot (left). Western blot assays determined the rescue effects of PRKCH on the expression of p53 and p21 in cotransfected cells (right). (C) qRT-PCR was used to determine the effects of PRKCH on p53 and p21 mRNA levels. (D) qRT-PCR detected the rescue effects of PRKCH on p53 and p21 protein expression in ACC-M cells. (E) ACC-M cells were cotransfected with pcDNA3-flag/p53 (Flag tag) and pA3M1/PRKCH (Myc tag) or pA3M1. The influence of PRKCH expression on the ubiquitination levels of p53 was determined by western blotting. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. All of the error bars indicate the means±SDs. All of the experiments were repeated at least three times.