Fig. 8

Myc affects MED1 occupancy at the TMEM44-AS1 super-enhancer in glioma cells. (A) Immunofluorescence staining to detect the co-localization Myc (red), and MED1 (green), and DAPI (blue) in LN-18 and U251 cells. (B) Co-IP assays were performed using LN-18 and U251 cells lysate with Myc or MED1 antibodies and detected by western blot with indicated antibodies. (C) Correlation between Myc and MED1 expression in the TCGA glioma dataset. (D-E) MED1 ChIP followed by qPCR analyzed the occupation of MED1 on the super-enhancer and promoter of TMEM44-AS1. The co-immunoprecipitated DNA was amplified by PCR using indicated primer. (F) The enrichment of MED1 on the enhancers of TMEM44-AS1 was analyzed with ChIP assays using the MED1 antibody after SF126 cells were transfected with vector or Myc plasmid. **P < 0.01 vs. vector. The independent biological experiments were repeated at least three times. (G-H) The luciferase reporter assays were used to detect the TMEM44-AS1 promoter, or constituent enhancers (E4 and E5) in LN-18 and U251 cells transfected with si-MED1 or NC. *P < 0.05, **P < 0.01 vs. NC. The independent biological experiments were repeated at least three times. (I) TMEM44-AS1 expression in LN-18 and U251 cells co-transfected with Myc plasmid and si-MED1. ^**p < 0.01 vs. vector; ^##P < 0.01 vs. Myc