Fig. 5
TRAF3IP2-AS1 accelerates the decay of PARP1 mRNA by recruitment of m6A methyltransferase complex. a The location of TRAF3IP2-AS1 binding site and a potential m6A site are indicated. Abundance of PARP1 transcript among mRNA immunoprecipitated with anti-m6A antibody was measured by qRT-PCR and normalized to IgG. b–c The mRNA level of PARP1 was detected by qRT-PCR after transfected with indicated vectors or siRNAs. d–e METTL3 was immunoprecipitated followed by qRT-PCR for assessing the association of the indicated PARP1 mRNA with METTL3 after overexpression or knockdown of TRAF3IP2-AS1. f–g Abundance of PARP1 among mRNA immunoprecipitated with anti-m6A antibody from cells transfected with indicated vectors or siRNA/ASO was measured by qRT-PCR. h PARP1-3′-UTR of the wild-type or containing a m6A consensus sequence mutant (A to G) was fused with a luciferase reporter. Luciferase activity of PARP1-3′-UTR was measured and normalized to Renilla luciferase activity. p–k Luciferase activity of PARP1-3′-UTR was measured after co-transfected with TRAF3IP3-AS1 and siMETTL3/siMEETTL14/siWTAP. l–m The stability of PARP1 mRNA in cells co-transfected with indicated vectors or siRNAs after treatment with α-amanitin. n Schematic illustration of targeted RNA methylation system. o–p Abundance of PARP1 among mRNA immunoprecipitated with anti-m6A antibody from cells transfected with indicated gRNA was measured by qRT-PCR. q The protein and mRNA level of PARP1 were measured by qRT-PCR and Western blot after transfected with indicated gRNA. r–s The stability of PARP1 and GAPDH mRNA in cells transfected with indicated gRNA after treatment with α-amanitin. The data are presented as the mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001