Fig. 4

AGAP2-AS1 epigenetically silences CDKN1A transcription by binding to EZH2. a CDKN1A expression was determined by qRT-PCR and Western blot assays after AGAP2-AS1 knockdown by transfection. b RIP assays were performed in HUCCT1and RBE cells and the coprecipitated RNA was subjected to qRT-PCR for AGAP2-AS1. c, d Bioinformatics predicts that there is a positive correlation between AGAP2-AS1 and EZH2 and a negative correlation between EZH2 and CDKN1A. e Western Blot assays were performed to determine the expression of EZH2 in HUCCT1 cells after EZH2 knockdown. f ChIP-qRT-PCR for EZH2 occupancy and H3K27me3 binding to the CDKN1A promoter in HUCCT1 and RBE cells. g, h CCK-8 assays were used to determine the viability of si-AGAP2-AS1 and si-CDKN1A co-transfected HUCCT1 cells. Representative images and data based on three independent experiments. Bars: ± sd, *P < 0.05, **P < 0.01