Fig. 5 | Nature Communications

Fig. 5

From: N6-methyladenosine modification of circNSUN2 facilitates cytoplasmic export and stabilizes HMGA2 to promote colorectal liver metastasis

Fig. 5

CircNSUN2 interacts with IGF2BP2 through CAUCAU motif. a Immunoblot analysis of IGF2BP2 after RNA-pulldown assay showing its specific association with circNSUN2. b RIP assays showing the association of IGF2BP2 with circNSUN2. Relative enrichment representing RNA levels associated with IGF2BP2 compared to an input control. IgG antibody served as a control. Data represent mean ± S.D. from three independent experiments; dot plot reflects data points from independent experiment. c IF-FISH assay showing that circNSUN2 is colocalized with IGF2BP2 protein in the cytoplasm. Scale bar, 10 µm. d Top, schematic structures showing RNA-binding domains within IGF2BP2 protein and a summary of IGF2BP2 truncations. Middle, relative enrichment representing circNSUN2 levels associated with truncated IGF2BP2 relative to an input control. Bottom, immunoblot analysis with anti-FLAG of HCT116 cells transfected with plasmids encoding FLAG-tagged WT or truncated IGF2BP2s. Data represent mean ± S.D. from three independent experiments; dot plot reflects data points from independent experiment. e Top, schematic illustration showing the CAUCAU motif located at exon 5−exon 4 junction site of circNSUN2 and the RNA probe for RNA-EMSA assay. Bottom, RNA-EMSA assay showing the binding ability of purified IGF2BP2 with biotin-labeled oligonucleotides containing CAUCAU motif from circNSUN2. Source data are provided as a Source Data file. Unprocessed original scans of blots are shown in Supplementary Fig. 10

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