FIGURE 4.
Direct regulation of the SESN2 expression by p53 and PAD4. A, RT-qPCR analyses of p53 and its target gene expression after 6 h of YW3-56 treatment of U2OS cells with or without p53 depletion by shRNA. The expression levels of each gene in control U2OS cells without p53 depletion and YW3-56 treatment were normalized to 1. Averages and standard deviations are shown (n = 3). B, representative Western blot results of the effects of YW3-56 on histone H3 citrullination and SESN2 expression in U2OS cells with or without PAD4 siRNA treatment. C, schematic illustration of the SESN2 gene intron and exon structure organization. Coordinates of the primers used to probe for p53 and PAD4 binding are indicated. TSS position is set as +1. D, ChIP analyses of p53 binding at the SESN2 gene before and after UV irradiation treatment for 6 h. E, ChIP analyses of p53 and PAD4 association as well as histone H3R17Me and H3Cit modifications at the +733 position of SESN2 gene before and after YW3-56 treatment at 6 μm for 6 h. Averages and standard deviations (n = 2 × 3) are shown (* indicates p < 0.005). TSS, transcription start site.