Extended Data Fig. 7: Functional analyses of C19MC in cell line models.
a) Scatter plot shows a re-analysis of the dataset published by Mong et al., 2020. C19MC is expressed in HEK293 cells using a CRISPR activation system and two different gRNAs specific for its promoter region. Downregulated genes (n = 509, putative C19MC targets) are located at the bottom left. C19MC targets identified by eCLIP in BT183 cells are indicated in red. Scatter plot at the bottom shows a magnified view of downregulated genes, with key C19MC targets labeled. b) Heatmap on the left shows relative gene expression of 75 downregulated genes that were also identified as C19MC targets by eCLIP. Heatmap on the right shows a detailed view for which C19MC member eCLIP peaks were detected. c) Brightfield images showing BT183 ETMR cell confluency after treatment for 72 h with LNAs targeting seven C19MC families, non-targeting CTRL-LNAs (CTRL1, CTRL2), or solvent control (H2O). Identified cell areas are highlighted in red. Scale bars = 400 µm. Images shown are representative of the n = 4 biological replicates. d) Brightfield and fluorescence images showing BT183 cell caspase 3/7 signal after treatment for 72 h with LNAs targeting seven C19MC families, non-targeting CTRL-LNAs (CTRL1, CTRL2), or Solvent control (H2O). Scale bars = 500 µm. Images shown are representative of the n = 4 biological replicates. e) Bar plots show quantification of protein expressing cells for the indicated markers assessed by immunofluorescence analysis. Percentages of positive cells from three replicates are shown as mean values with SEM. P-values were calculated using the two-sided, unpaired Student’s t-test. Related to Figs. 4 and 5.
