Fig. 6
Myc bind to TMEM44-AS1 super-enhancer and promoter region, and promoted its transcription. (A) RT-qPCR assays measuring the expression of TMEM44-AS1 upon knockdown of Myc in LN-18 and U251 cells. *P < 0.05, **P < 0.01. (B) Expression of TMEM44-AS1 in SF126 cells was examined transfected with vector or Myc plasmid. **P < 0.01. (C) ChIP-seq profiles of H3K27ac, Myc, MED1, and RNA pol II in GBM cells and H3K27ac in normal brain tissues. (D-E) Myc ChIP followed by RT-qPCR analyzed the occupation of Myc on the super-enhancer and promoter of TMEM44-AS1. The co-immunoprecipitated DNA was amplified by PCR using indicated primer. (F-G) Enhancer and promoter activity measured by luciferase reporter assays in LN-18 and U251 cells. Three constituent enhancers (E1, E4 and E5) within the SE were separately cloned into luciferase reporter vector pGL3-enhancer and the promoter was cloned into luciferase reporter vector pGL3-promoter. (H) Luciferase assay analysis of SF126 cells transfected with TMEM44-AS1 promoter, or constituent enhancers (E1, E4 and E5) constructs together with Myc or vector