Fig. 10

Drug-tolerant cells exhibit an altered differentiation state that is also present in human medulloblastoma. a Model for altered differentiation in BETi-tolerant medulloblastoma cells. Human medulloblastomas express both stem-cell and neuronal differentiation markers. Following treatment with BETi, drug-tolerant cells exhibit a more neuronal phenotype however do not undergo terminal differentiation, maintaining the ability to progress through the cell-cycle. b Expression levels of TUJ1 (TUBB3) and JQ1 sensitivity of 46 central nervous system cell lines from the CTRP database. AUC denotes area under the curve. Asterisks denote statistically significant differences (*p < 0.05, **p < 0.01, ***p < 0.001) as determined by a two-tailed unpaired t-test. Source data: Source Data File. c Representative images of three medulloblastomas stained for TUJ1 (green), SOX2 (red), NESTIN (yellow), OLIG2 (white) and DAPI (blue). Scale bar is 100 μm). Source data: Supplementary Data File 9. d Percentage of D458 sensitive and drug-tolerant cells that are TUJ1 positive and MSI1 negative in the presence of JQ1 (1 μM) or DMSO control. Data from three independent experiments are shown; error bars depict mean ± SEM. Asterisks denote statistically significant differences (*p < 0.05, **p < 0.01, ***p < 0.001) as determined by a two-tailed unpaired t-test. Source data: Supplementary Data File 9. e Schematic depicting barcoding experiments with 600,000 barcode library. A pooled lentiviral library encompassing 600,000 unique DNA barcodes was used to label individual cells. Cells were expanded before being placed in replicate DMSO or JQ1 drug-treatment experiments. Barcodes were PCR amplified prior to treatment (ETP = early time point) and from replicate experiments at the drug of treatment and subjected to next-generation sequencing. f Percentage of shared DNA barcodes across replicates of barcoded D458 cells treated with JQ1 (2 μM). Within each replicate, the number of other replicates with whom each barcode is shared is depicted. Source data: Source Data File