Fig. 4

TRAF3IP2-AS1 down-regulates PARP1 mRNA by direct binding. a The RNA levels of TRAF3IP2-AS1 and PARP1 were detected after transfection with TRAF3IP2-AS1 plasmids or siTRAF3IP2-AS1 for 48 h. b Level of PARP1 mRNA detected by qRT-PCR after MS2-RIP for GFP in UOK109 cells. AS1, NC, AS1-antisense and AS1-Mut correspond to TRAF3IP2-AS1, empty vector, TRAF3IP2-AS1-antisense and TRAF3IP2-AS1 with mutation of potential binding site to PARP1 mRNA, respectively. c–d Dual Luciferase Reporter Assay used to detect the relative luciferase activity in HEK293 T cells co-transfected with siTRAF3IP2-AS1 and pmirGLO-PARP1 3′-UTR WT/MUT. e The stability of PARP1 mRNA and GAPDH mRNA in UOK109 cells transfected with TRAF3IP2-AS1 or TRAF3IP2-AS1 contained binding site mutation was measured by qRT-PCR relative to 0 h after blocking new RNA synthesis with α-amanitin and normalized to 18 s rRNA. f The RNA levels of NONO-TFE3 and PARP1 were detected after transfection with NONO-TFE3 plasmids or shTFE3 for 48 h. g The stability of PARP1 mRNA and GAPDH mRNA in 786-O cells transfected with siTRAF3IP2-AS1 was measured by qRT-PCR after treatment with α-amanitin. h The protein level of PARP1 was detected after transfection with TRAF3IP2-AS1 plasmids or siTRAF3IP2-AS1