Fig. 5: IGF2BP2 interacts with CENPB mRNA and regulates its expression.

a Localization of CENPB mRNA in HT29 cells, as revealed by smFISH staining of CENPB (green) and nuclei (blue). b Identified IGF2BP2-binding sequence in the 3′UTR and CDS regions of CENPB mRNA. Top, genomics locus and diagram of CENPB mRNA. Bottom, IGF2BP2 binding peaks and wiggle-tracks of data from eCLIP-seq in K562 cells. LNAs pairing to CENPB-3′UTR are shown as navy lines. c Schematic of in-cell and in vitro RNA pull-down assay. d, e CENPB-3′UTR, but not CENPB-CDS, preferentially to bind to IGF2BP2, as revealed by in-cell (top panel) and in vitro (bottom panel) RNA pull-down assay (d) and EMSA assay (e). f IGF2BP2 enriched more endogenous CENPB mRNA, as detected by native RIP assay. IgG was performed as a negative control; GAPDH mRNA and U6 RNA were detected as negative controls. g LNA-blocked RNA pull-down assay revealed the specific binding sites of IGF2BP2 on CENPB-3′UTR. h, i KD of IGF2BP2 (h) and CWI1-2 (5 nM and 10 nM) treatment (i) reduced CENPB expression in DLD-1 cells, as shown by RT-qPCR (left panel) and WB (right panel). j LNA-b2 reduced CENPB expression in DLD-1 cells. Images are representative of three independent experiments (a, d, e, g–i). RNA levels were detected by RT-qPCR (f, h–j), normalized to ACTB (h–j) and are presented as mean ± s.d., with biologically individual data points shown (f, h–j). P values were determined by ordinary two-way ANOVA with Dunnett’s post-hoc comparisons versus scramble control at each row factor level (adjusted P values) (f), ordinary one-way ANOVA test with Dunnett’s post-hoc comparisons versus scramble control (FDR-adjusted P values) (h, i), and unpaired two-tailed Student’s t-test (j). Source data are provided as a Source Data file.