Fig. 1: CREB5 regulates ID1 expression independently of the BMP signaling pathway in H3.3K27M DIPG.

a H&E-stained sections or immunohistochemistry for H3K27M, ID1, and H3K27me3 in human non-DIPG pontine and DIPG samples. Scale bar: 50 μm. b Immunoblotting analysis of the indicated proteins in PPCs, the H3.1K27M DIPG cells (DIPG-IV), and the H3.3K27M DIPG cells (DIPG17 and TT150630). c The bioluminescence activity (left) and representative bioluminescence images (BLI) (right) in TT150630 mouse models between ID1 KD and Ctr (n = 5). d Kaplan-Meier survival analysis from animals implanted with TT150630 cells with or without ID1 KD in the pons. e Immunofluorescence images of pons sections from TT150630 PDX animals with or without ID1 KD for anti-OLIG2, anti-GFAP, and anti-β-tubulin with quantification. Scale bar: 20 μm. Right panel: quantified cell counts. f Immunoblotting analysis of indicated proteins in DIPG17 cells with or without ID1 KD. g ID1 expression across indicated DIPG subtypes from PedcBioPortal and UCSC Xena DIPG cohort. h, Immunoblotting analysis (top) and quantification (bottom) of ID1 expression in indicated DIPG cells treated with LDN-193189 (200 nM) for 0, 2, 4, and 8 hours. i Clustering heatmap showing regulon specificities in OPC-like cells. j qPCR analysis of ID1 expression in indicated DIPG cells. k Kaplan-Meier survival curves for DIPG patients from UCSC Xena cohort separated into CREB5 high and low expression groups. l, Dual-luciferase reporter assay using ID1 promoter. m qPCR analysis of CREB5 and ID1 in indicated DIPG cells. n Scatter plot showing the correlation of ID1 and CREB5 expression in clinical pediatric brain tumors (CBTTC). The p value and correlation coefficient (R) were calculated using Pearson’s correlation. The error bands indicate 95% CIs as shades based on standard error. Data presented as mean ± s.e.m. of three independent experiments (e, j, l, and m) or mean ± s.e.m. (c); statistical significance was determined by a two-tailed unpaired Student’s t-test (c, e, j, l, m, and n), log-rank test (d and k) or ordinary one-way ANOVA with Tukey test (g and h). Experiments were repeated three times independently with similar results (b, f and h). Source data are provided as a Source Data file.