Fig. 1: OLIG2 expression in pediatric MB and OLIG2 siRNA knockdown studies.

A Kaplan-Meier curves of the SHH-driven MBα patients based on OLIG2 expression. B OLIG2 expression in PDX models, primary cell lines and ATCC pediatric MB cell lines (data are shown as means ± SD, n = 6). C OLIG2 protein expression in MB cell lines. D H&E and IHC for OLIG2 and Ku80 in MB Med-813 xenografts. OLIG2+ and Ku80+ cells shown in white. Scale bars as indicated. E Representative western blots show OLIG2 expression in Daoy cells 72 hours after indicated siRNA. F Representative western blots show cC3 expression, total caspase-3, cleaved PARP and total PARP in Daoy cells 72 hours after indicated siRNA. Three different blots were used, identified by no asterisk, * and **, and a separate β-actin loading control is shown for each blot. G Representative western blots show expression of cyclin B1, p-CDK1, p-HH3, total CDK1 and total HH3 in Daoy cells 72 hours after indicated siRNA. Three different blots were used, identified by no asterisk, * and **, and a separate β-actin loading control is shown for each blot. H Daoy cells stalled in G₂/M phase 72 hours after siRNA OLIG2 knockdown (means ± SD, n = 6, p value by 1-way ANOVA). I Top panel shows a Daoy cell in anaphase with normal nuclear morphology and spindle alignment. siRNA OLIG2 knockdown results in abnormal nuclear phenotypes including satellite micronuclei. Scale bars are indicated in I. J Bright field images of Daoy cells transfected with scrambled control sequence and siRNA OLIG2 KD Seq #1 over 48 hours. Cells labelled with cleaved Caspase-3/7 were shown in green. K Representative study showing cell confluence (left) and cleaved caspase-3 confluence (right) of Daoy after siRNA knockdown compared to scrambled control, wild-type Daoy with or without lipofectamine. Graph shows mean and standard deviation of 16 measurements (technical replicates) within individual wells. The study was repeated two additional times with similar values, included in Supplementary Data 4 Source Data File. B, H dots indicate individual biological replicates and columns indicate means. Source data are provided as Supplementary Data 4 Source Data file.