Fig. 4

ZFPM2-AS1 upregulated TRAF4 expression via sequestering miR-3612. a Three downstream genes (TRAF4, CBX5, TAOK1) were predicted as potential targets of miR-3612. b qRT-PCR and western blot assays measured the mRNA and protein levels of TRAF4, CBX5 and TAOK1 in SHP-77 and DMS-53 cells. c, d Luciferase activity of TRAF4-WT or TRAF4-MUT was examined by luciferase reporter assay in SHP-77 and DMS-53 cells. e RIP assay tested the enrichment of TRAF4, miR-3612 and ZFPM2-AS1 in Anti-Ago2 group. f RNA pull down assay measured the enrichment of TRAF4 and ZFPM2-AS1 in Bio-miR-3612 sense group. g, h The mRNA and protein expressions of TRAF4 in differently transfected groups were assessed by qRT-PCR and western blot. *P < 0.05, **P < 0.01, *** P < 0.001