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. 2021 Aug 30;12(9):816. doi: 10.1038/s41419-021-04014-x

Fig. 5. STING is required for 2′3′-cGAMP/BV6/zVAD.fmk-induced cell death.

Fig. 5

A Western blot analysis of STING expression in the indicated PC cell lines after 48 h of transfection with non-silencing RNA (ctrl) or siRNA targeting STING. GAPDH serves as loading control. Representative blots of at least two different independent experiments are shown. B AsPc-1 and BxPc-3 cells were transfected with control siRNA (siCtrl) or two independent siRNAs targeting STING and were treated with 20 μM zVAD.fmk for 48 h in the presence or absence of 4 µg/ml 2′3′-cGAMP and 5 µM BV6. The amount of cell death was calculated by quantifying PI uptake determined with the ImageXpress Micro XLS system. Data are presented as percentage of PI-positive cells, and mean and SD of three independent experiments performed in triplicate are shown. **P < 0.01, ***P < 0.001. C Western blot analysis of AsPc-1 and BxPc-3 subjected to nHT and STING CRISPR/Cas9 knockdown. β-Actin serves as loading control. Representative blots of at least two different independent experiments are shown. D AsPc-1 and BxPc-3 cells were subjected to nHT and STING CRISPR/Cas9 knockdown, and treated with 20 μM zVAD.fmk for 48 h in the presence or absence of 4 µg/ml 2′3′-cGAMP and 5 µM BV6. The amount of cell death was calculated by quantifying PI uptake determined with the ImageXpress Micro XLS system. Data are presented as percentage of PI-positive cells, and mean and SD of three independent experiments performed in triplicate are shown. ***P < 0.001.