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. 2024 Oct 22;10(21):e39677. doi: 10.1016/j.heliyon.2024.e39677

Fig. 1.

Fig. 1

Confirmation of PTPN11 as an essential gene for OSCC survival. (A) Plot of number of dependent OSCC cell lines based on our previous CRISPR/Cas9 screen of 21 OSCC lines (Chai et al., 2020), for the essential genes that are enriched in the lineage of upper aerodigestive tract (DepMap Public 22Q4, https://depmap.org/portal/). (B) Scatter plots of CRISPR scores of PTPN11 and other genes that are targets of drugs being tested for OSCC. (C) Co-competition assays between non-transduced cells with either Non-targeting (NT) or PTPN11 sgRNAs-transduced OSCC to validate dependency on PTPN11. The percentage of BFP-positive transduced cells obtained at different time points were normalized to the day 4 or 6 readings for respective sgRNAs. (D) Western blots showing depletion of the SHP2 level, the protein encoded by PTPN11 gene upon sgRNA-mediated knockout. Actin is used as loading control. NT – Non-targeting sgRNAs; “P1k” – PTPN11_sg1k”; “P2b” – PTPN11_sg2b. (E) Cell viability assay of OSCC cells upon PTPN11 knockout with P1k and P2b. (F) Colony forming assay of OSCC cells upon PTPN11 knockout with P1k and P2b. (G) Apoptosis assay of OSCC cells upon PTPN11 knockout with P1k and P2b.