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. 2019 Dec 5;27(6):1782–1794. doi: 10.1038/s41418-019-0461-z

Fig. 7. IGF2BP2 regulates DANCR mRNA stability through m6A modification at A664.

Fig. 7

a IGF2BP2 interacts with DANCR, as determined by RIP assay using IGF2BP2 antibody. b Detection of in vivo interaction between IGF2BP2 and DANCR by S1m tagged DANCR pulldown. In vivo expressed S1m tagged DANCR was first pulled down by streptavidin beads and IGF2BP2 was then detected by western blot. c IGF2BP2 siRNA reduces DANCR stability in BXPC-3 cells. Cells were first transfected with IGF2BP2 siRNA#3 and control siRNA. Then, the transfected cells were treated with 2 µg/ml actinomycin D and total RNA was isolated at various time points as indicated. d IGF2BP2 KO also reduces DANCR stability. e Enrichment of DANCR by RIP assay with m6A antibody. f Detection of interaction of DANCR with m6A antibody. In vivo expressed S1m tagged DANCR was first pulled down by streptavidin beads and then incubated with m6A antibody. The signals bound to the membrane were detected by donkey anti-mouse antibody conjugated with IRDye® 680RD. HL heavy chain, LC light chain. g Mutation A664 in DANCR abolishes its interaction with m6A antibody. WT wild type, Mut mutant. Values in a, c, d, e are mean ± SEM. **P < 0.01; ***P < 0.001.