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Fig. 4 | Clinical Epigenetics

Fig. 4

From: CRISPR-Cas9 knockout screen identifies novel treatment targets in childhood high-grade glioma

Fig. 4

Successful validation in CSC. A Thirteen genes were individually knocked in GU-pBT-7 and GU-pBT-19 (n = 2 gRNA per gene, i.e. biological replicates, except negative control where n = 3 or 4 samples) and cultured for one month. The cell counts at each split was summarised and used to compare the growth of the knockout cells to the negative controls. All genes significantly decreased (Welch t test, p value < 0.05; indicated with * above the bar) the CSC growth after knockout compared to the control for at least one of the cell lines. PLK1 was included as a positive control. B The six genes that showed the most effect were selected for further validation in two additional CSC lines and tested in two NSC lines (n = 2 gRNA per gene). Note that the data for GU-pBT-7 and GU-pBT-19 are from the same experiment as presented in the A-panel. Error bars represent the standard error mean. * above a bar indicates significant decrease in cell number for that gene knockout compared to the negative control for the cell line in question. # (with associated braces indicating the CSC and NSC lines) denotes a significant difference at group level between the CSC and NSC in the case of UBE2N, LSM11 and CHD4 knockout, respectively (Welch t test, p value < 0.05). C CHD4 knocked out in the CSC GU-pBT-19 showed drastic reduction in cell viability, but the NSC FT3477 were not affected morphologically, which was also seen for knockout of D UBE2N and E LSM11. The cell images are from around 10 days after knockout. Scale bar = 200 µm applies to all images in the C-E panel

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