Fig. 2

m6A sites were identified in NEAT1–1 in prostate cancer. a m6A RIP-seq analyswas of m6A sites of NEAT1 by two independent antibodies. The m6A profiles of NEAT1 were shown in genome browser. b Quantitative PCR verification of m6A RIP results on the gene body of NEAT1–1with m6A antibody against the m6A sites in P-18 primary cells. Means and standard deviations (error bar) were determined from three replicates. Error bars represent mean ± SD for triplicate experiments. P values were shown in the figures. c The top hits of NEAT1–1 pull-down proteins were identified by TAP-MS. The numbers of peptides were indicated in the lwast. d Ontology analyswas were by DAVID (david.ncifcrf.gov) and KEGG analyswas by DAVID and KAAS (www.genome.jp/kass-bin). e Biotin pull-down assay by incubating biotin-labeled specific probes targeting NEAT1–1 with P-18 cell lysate followed by Western blot with CYCLINC1, CYCLINL1 and CYCLINT1 antibodies. f CLIP-qPCR analyswas of CYCLINL1 binding at the NEAT1–1 in P-18 control or METTL3 knocking out cells transfected with control or METTL3-specific sgRNA. Immunoprecipitated RNAs were detected by real-time PCR. All data shown were mean values ± SD (error bar) from three replicates. P values were shown in the figures. g NEAT1–1 pull-down assay use biotin-labeled NEAT1–1 and NEAT1–1 mutants (mut). The RNA and protein complexes were detected by western blot