Fig. 5
NKAP is a reader of m6A on pri-miR-25. a The NKAP-binding sites on pri-miR-25 detected by CIMS and CITS miCLIP-seq. Orange tracks of NKAP-iCLIP-seq are unique tag coverage and blue tracks represent C>T transition and truncation, respectively. Filled purple circles denote iCLIP-called NKAP-binding site and the horizontal blue bars indicate the transcript model of pre-miR-25. b The DGCR8-binding sites on pri-miR-25 detected by DGCR8 HITS-CLIP. Filled purple circles denote iCLIP-called NKAP-binding site and the horizontal blue and red bars represent the transcript model of pre-miR-25 and the DGCR8-binding site, respectively. c In vivo binding of NKAP-FLAG to pri-miR-25 in PDAC cells exposed to CSC or DMSO detected by CLIP-qPCR. Values are the mean ± S.D. from three independent experiments. d The levels of NKAP-FLAG bound to pri-miR-25 were significantly elevated by overexpression of METTL3, but significantly decreased by knockdown of METTL3 in both PANC-1 (left panel) and BXPC-3 (right panel) cells. Values are the mean ± S.D. from three independent experiments. e Data of m6A residues and NKAP-binding sites identified by the analysis of cross-linking-induced mutation sites (CIMS) or cross-linking-induced truncation sites (CITS) in iCLIP-seq data. f Venn diagram showing overlap between m6A residues and NKAP-binding sites. g Significant enrichment of RGAC motif is in NKAP footprints (CIMS or CITS along with 5-nucleotide flanking sequences) analyzed by MEME SUIT with default parameters. h Shown is the intensity of NKAP binding centered at m6A residues. i Shown is the intensity of m6A CLIP signal centered at NKAP-binding sites. j Shown is the distribution of NKAP iCLIP tag density around the splicing factor binding sites. k Shown is the binding intensity of NKAP at splicing sites near or not near m6A residues. All statistic examinations in this figure are Student t test unless specific. *P < 0.05, **P < 0.01, and ***P < 0.001 compared with the corresponding control
