Figure 1 | Scientific Reports

Figure 1

From: CRISPR activation screen identifies TGFβ-associated PEG10 as a crucial tumor suppressor in Ewing sarcoma

Figure 1

Establishment of the CRISPRa cell line. (A) Schematic representation of the modified CRISPRa Synergistic Activation Mediator (SAM) system (Based on 22). (B) Schematic representation of the clonal cell line generation. (C) Expression of the surface marker CD44 was measured by flow cytometry after introduction of CD44 targeting gRNAs. The percentage of positive cells for three gRNAs and four cell clones (A3, A4, H5 and D8) is indicated. Image was created by using FlowJo version 10, https://www.flowjo.com. (D) mRNA expression of four repressed genes (CD44, LOX, PHLDA1, and IGFBP3) was measured by qRT-PCR after gRNA transduction in the clones A3 and D8. Data is indicated as fold change over non-targeting control guide AAVS1, normalized to GAPDH. Image was created by using GraphPad version 8, https://www.graphpad.com. (E) Schematic representation of the gene selection for the lentiviral library LIBerty.

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