Figure 2
RFX5 binds to the promoter region and activates transcription of the KDM4A genes in HCC. (A) Candidate RFX5 target genes determined by analyzing the RFX5 ChIP-seq binding peak and signal of HepG2 cells from ECODE dataset and RNA-seq data from TCGA LIHC dataset. (B) The ChIP-seq binding peak and signal of RFX5 in KDM4A promoter regions. The histone modification status and the amplification region of ChIP-PCR were also indicated. (C) The immunoprecipitated DNA fragments produced using anti-RFX5 antibody underwent PCR analysis of KDM4A promoter region in HCC cells. (D) The transcriptional activity of KDM4A promoter reporter constructs (pGL4-KDM4A) was measured in MHCC-97H utilizing relative luciferase activity (RLA), which was the ratio of firefly to Renilla luciferase activities. The effect of modulation of RFX5 expression on KDM4A transcriptional activity was determined by co-transfection of pGL4-KDM4A and FLAG-RFX5 in MHCC-97H cells. (E) The effect of RFX5 expression on the transcriptional activity of KDM4A was identified through co-transfection of sgRNAs targeting RFX5 (RFsg1 or RFsg3) and pGL4-KDM4A in MHCC-97H cells. *P < 0.05, **P < 0.01, ***P < 0.001.
