Figure 5
PEG10 expression affects cell proliferation in a dose-dependent manner and is repressed in the presence of EWS-FLI1. (A) Schematic representation of the inducible dCas9-VP64 construct. (B) PEG10 protein level in SKNMC cells with the inducible CRISPRa system overexpressing PEG10 that were harvested 4 days post induction. Image was created by using Image Lab version 6.1, https://www.bio-rad.com/en-ch/product/image-lab-software. (C) Quantification of the number of SKNMC cells with the inducible CRISPRa system overexpressing PEG10 over 9 days post induction (n = 2,* p < 0.1, **p < 0.01, ***p < 0.001, paired t-test). Image was created by using GraphPad version 8, https://www.graphpad.com. (D) EWS-FLI1, PEG10 and SGCE protein levels in SKNMC cells with the inducible EWS-FLI1 shRNA construct upon induction for 24, 48 and 72 h. Data is representative for 3 biological replicates. Image was created by using Image Lab version 6.1, https://www.bio-rad.com/en-ch/product/image-lab-software. (E) PEG10 mRNA expression levels across 15 EWS-FLI1 positive Ewing sarcoma cell lines after EWS-FLI knockdown via inducible EWS-FLI1 shRNA for 96 h. Data was extracted from the Ewing Sarcoma Cell Line Atlas (51) (n = 3,* p < 0.1, **p < 0.01, ***p < 0.001, paired t-test). Image was created by using GraphPad version 8, https://www.graphpad.com.
