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. 2020 Dec 1;10(12):4287–4307.

Figure 3.

Figure 3

Cellular cisplatin tolerance was increased with downregulation of GSDME expression and vice versa. A. Immunoblot confirmed that GSDME was overexpressed in NOKs and GSEs; the Flag-tag was successfully fused to GSDME; B. The cytotoxicity assay revealed elevated LDH release in GSDME-overexpressing cells compared with control cells after cisplatin treatment, suggesting more GSDME-ov cell death at the same cisplatin concentration; C. Immunoblot analysis confirmed that GSDME was knocked out by GSDME sgRNA in NOKs and GSEs; D. The cytotoxicity assay revealed decreases in LDH release in GSDME knockout cells compared with control cells after cisplatin treatment; E. Gradient concentration of cisplatin-treated GSDME-overexpressing cell lines; cisplatin tolerance was decreased in NOKs and GSEs; F. Gradient concentration of cisplatin-treated NOKs and GSEs with GSDME knockout group; increased cisplatin tolerance was detected; G. FCM detected the cisplatin-sensitive effect on GSDME-overexpressing cell lines using NOKs and GSEs; H. FCM detected the cisplatin-tolerant effect on the GSDME knockout cell line using NOKs and GSEs; I. Statistical analysis of cell death rates of G; cisplatin induced more cell death in GSDME-overexpressing cell lines; J. Statistical analysis of cell death rates of H; GSDME knockout cell lines were more tolerant during cisplatin treatment. (cisplatin treatment: NOK 30 μM; GSE 20 μM; HUVEC 40 μM, 24 hours for B, D, G, H).