Fig. 3

MYBL2 mediates its phenotype via direct upregulation of CCNF, BIRC5, and AURKB. a RNA-seq showing differentially expressed genes (DEGs) after siRNA-mediated MYBL2 knockdown compared to a non-targeting siControl. A summary of three cell lines is shown; n = 3 technical replicates per condition. b GSEA of RNA-seq data. Displayed are 275 gene-sets downregulated upon MYBL2 knockdown that had an FDR q < 0.05. c Analysis of MYBL2 ChIP-seq data from A673 cells showing MYBL2 peaks in the promoters of CCNF, BIRC5, and AURKB. Publicly available EWSR1-FLI1 ChIP-seq data from A673 cells was analyzed to exclude a direct regulation by EWSR1-FLI1. Whole-cell extract (WCE) served as a control. d Linear regression of CCNF, BIRC5, and AURKB expression onto MYBL2 expression in 166 EwS tumors. e Kaplan–Meier survival analyses of 166 EwS patients stratified by median expression levels of the indicated gene; P values determined via Mantel–Haenszel test. f Viable cell count 96 h after transfection of A673 and SK-N-MC cell lines with two different siRNAs directed against either CCNF, BIRC5, or AURKB (summary of two different siRNAs shown) or a non-targeting siControl. Horizontal bars represent means, and whiskers represent the SEM, n ≥ 3 biologically independent experiments. g Measurement of cell death using Trypan blue positivity 96 h after transfection of A673 and SK-N-MC cells transfected as described in f. Horizontal bars represent means, and whiskers represent the SEM, n ≥ 3 biologically independent experiments. ***P < 0.001, **P < 0.01, *P < 0.05; P values determined via two-tailed Mann–Whitney test. Source data are provided as a Source Data file