Fig. 5: Regulation of IGF2BP3 on m7G-marked transcripts in glioblastoma cells.
From: IGF2BP3 promotes mRNA degradation through internal m7G modification
a Gene Set Enrichment Analysis (GSEA) of genes in ‘Glioma’ KEGG pathway against ranked list of genes according to half lifetime changes upon IGF2BP3 knockdown. b Kaplan-Meier survival analysis in TCGA database for Glioma (TCGA-GBMLGG) with wildtype TP53 (left), or with mutant TP53 (right). The patients were divided into two groups of equal size based on IGF2BP3 levels. P-value detected by log-rank test. c Changes in TP53 mRNA level in highly methylated cells upon IGF2BP3 knockdown. Mean ± SEM (n = 3) with two-tailed Student’s t-tests. Calculated half lifetimes marked in corresponding colors. d–f Relative m7G levels of the TP53 3’ UTR locus upon METTL1 knockdown (d), IGF2BP3 overexpression (e), and IGF2BP3 knockdown (f), normalized to the methylation level in the corresponding control cells. Mean ± SEM (n = 3) with two-tailed Student’s t-tests. g Changes in mRNA levels in cells with introduction of dCas13b-IGF2BP3 or dCas13b-METTL1 and TP53 gRNA at the loci or not (neg). Mean ± SEM (n = 3) with two-tailed Student’s t-tests. Calculated half lifetimes marked in corresponding colors. h Relative m7G levels of the TP53 3’ UTR locus with introduction of dCas13b-IGF2BP3 or dCas13b-METTL1 and TP53 gRNA at the loci or not (neg), normalized to the methylation level in wildtype cells. Mean ± SEM (n = 3) with two-tailed Student’s t-tests. i Relative TP53 mRNA levels in cells with dCas13b-IGF2BP3 or dCas13b-METTL1 and the guide RNA, normalized to the cells with negative gRNA. Mean ± SEM (n = 3) with two-tailed Student’s t-tests. j Western blot of p53 levels in T98G cells with dCas13b-IGF2BP3 or dCas13b-METTL1 tethering with the guide RNA or the negative gRNA. k Cell proliferation in LN229 cells with dCas13b-IGF2BP3 tethering with the guide RNA or the negative gRNA. Mean ± SEM (n = 6) with two-tailed Student’s t-tests. l Western blot of MGMT levels in T98G cells with dCas13b-IGF2BP3 tethering with the guide RNA compared to negative gRNA and wildtype cells. m Survival percentages of T98G cells treated TMZ at different concentrations with dCas13b-IGF2BP3 tethering with the guide RNA, or the negative gRNA, compared to wildtype cells. Mean ± SEM (n = 6) with two-tailed Student’s t-tests. j and (l) were both repeated independently twice with similar results. All source data are provided as a Source Data file.
