Research Paper Volume 13, Issue 9 pp 13179—13194

C/EBPα is indispensable for PML/RARα-mediated suppression of long non-coding RNA NEAT1 in acute promyelocytic leukemia cells

C/EBPα directly binds and transactivates the promoter region of NEAT1. (A) Upper panel: Schematic representation of putative C/EBP binding sites in the NEAT1 promoter. Lower panel: C/EBPα ChIP-qPCR showing the enrichment of C/EBPα in each putative binding site, the negative control and positive control (SPI1 promoter) in NB4 cells that were untreated or treated with ATRA at 1μM for 24 h (RA 24h). (B) ChIP was performed on two APL patient samples with anti-C/EBPα antibody. DNA fragments at NEAT1 promoter were subsequently measured with qPCR. (C) The 1656 bp NEAT1 promoter reporter construct (125 ng) was transfected into 293T cells along with pcDNA3.1 vector (empty) or pcDNA3.1-C/EBPα (C/EBPα) expression plasmid (500 ng). The data represent the mean ± S.E.M from 3 replicates.

Figure 1. C/EBPα directly binds and transactivates the promoter region of NEAT1. (A) Upper panel: Schematic representation of putative C/EBP binding sites in the NEAT1 promoter. Lower panel: C/EBPα ChIP-qPCR showing the enrichment of C/EBPα in each putative binding site, the negative control and positive control (SPI1 promoter) in NB4 cells that were untreated or treated with ATRA at 1μM for 24 h (RA 24h). (B) ChIP was performed on two APL patient samples with anti-C/EBPα antibody. DNA fragments at NEAT1 promoter were subsequently measured with qPCR. (C) The 1656 bp NEAT1 promoter reporter construct (125 ng) was transfected into 293T cells along with pcDNA3.1 vector (empty) or pcDNA3.1-C/EBPα (C/EBPα) expression plasmid (500 ng). The data represent the mean ± S.E.M from 3 replicates.