Fig. 3

IGF2BP2-mediated m⁶A modification favors the stability of TRPM2-AS. A m⁶A modification sites of TRPM2-AS were predicted using SRAMP (http://www.cuilab.cn/sramp). B MeRIP and RT-qPCR assays revealing the m⁶A enrichment of TRPM2-AS by using three different primers. C The relative expression levels of TRPM2-AS in GBC cells were measured by RT-qPCR with/without METTL3/METTL14 overexpression. D RT-qPCR showing the decreased expression of TRPM2-AS upon treatment with 0, 50, 100, 200 μM DAA. E RIP analysis revealing the binding levels of TRPM2-AS with IGF2BP1, IGF2BP2, IGF2BP3, YTHDF1and YTHDF3. F The binding of TRPM2-AS to IGF2BP2 was identified using a dual-luciferase reporter system. G Western blot validation of IGF2BP2 protein expression levels in IGF2BP2 stable overexpression/knockdown GBC cells by lentivirus transfection. H Relative expression level of TRPM2-AS in IGF2BP2 overexpression/knockdown GBC-SD and NOZ cells after treated with actinomycin for different time point. I RT-qPCR showing the relative expression level of TRPM2-AS with IGF2BP2 overexpression/knockdown. J RIP analysis revealing the binding of TRPM2-AS and IGF2BP2 after the m⁶A methylation of TRPM2-AS was inhibited by DAA. K S1m pull-down assessment showing the changes in IGF2BP2 level with three different TRPM2-AS m⁶A site mutations. L RIP analysis verifying the enrichment of TRPM2-AS in the control group and in the IGF2BP2 overexpression group with transfection of TRPM2-AS or TRPM2-AS m⁶A sites mutation (Mut-1) overexpression plasmid. Data were assessed with unpaired Student’s t test or one-way ANOVA and presented as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001