Fig. 4

DUXAP8 epigenetically silences CDKN1A and KLF2 transcription by binding with EZH2. a, b The levels of CDKN1A, CDKN2B, KLF2, Bcl-2, Bax, Caspase-3, Caspase-9, PTEN and EMP1 mRNA were detected by qPCR upon knockdown of DUXAP8 in BxPC-3 and PANC-1 cells. c Bioinformatic analysis to predict the interaction probabilities of DUXAP8 and RNA binding proteins via RNA–protein interaction prediction (http://pridb.gdcb.iastate.edu/RPISeq/). Predictions with probabilities > 0.5 were considered positive. RPISeq predictions are based on Random Forest (RF) or Support Vector Machine (SVM). d. RIP assays were performed in BxPC-3 cells and the coprecipitated RNA was subjected to qPCR for DUXAP8. e, f LSD1 and EZH2 expression levels were determined by qPCR when LSD1 or EZH2 were knocked down in BxPC-3 cells. g, h The levels of CDKN1A, CDKN2B, KLF2, Bcl-2, Bax, Caspase-3, Caspase-9, PTEN and EMP1 mRNA after knockdown of LSD1 or EZH2 in BxPC-3 cells. i Overlap of up-regulated genes after knockdown of DUXAP8, LSD1 and EZH2. j ChIP-qPCR of H3K4me2 and LSD1, H3K27me3 and EZH2 of the promoter region of the CDKN1A locus after siRNA treatment targeting si-NC or si-DUXAP8 in BxPC-3 cells. k ChIP-qPCR of H3K4me2 and LSD1, H3K27me3 and EZH2 of the promoter region of the KLF2 locus after siRNA treatment targeting si-NC or si-DUXAP8 in BxPC-3 cells. Bars: s.d, *P < 0.05, **P < 0.01