Osthole Induces Apoptosis and Caspase-3/GSDME-Dependent Pyroptosis via NQO1-Mediated ROS Generation in HeLa Cells

Figure 2

Osthole induces GSDME-mediated pyroptosis in HeLa cells. (a) After treatment with osthole, the levels of apoptosis-related protein were determined by western blotting. (b) The intensity of bands was quantified by Image J, and β-actin was used as a control. (c–e) Pretreatment with 20 μM Z-VAD-FMK for 1 h prior to treatment with 320 μM osthole for 18 h. (c) Cell viability was detected by MTT assay. (d) Cell death was determined using Annexin V-FITC/PI double staining by flow cytometry. (e) Percentage (%) of cells stained by Annexin V-FITC and/or PI. (f) The expression of GSDME and GSDME-N was analyzed by western blotting at protein levels. (g) The intensity of bands was quantified by Image J, and β-actin was used as a control. (h–l) GSDME was knocked down in HeLa cells. (h) Cell viability was detected by MTT assay. (i) Cell death was detected using Annexin V-FITC/PI staining by flow cytometry. (j) Percentage (%) of cells stained by Annexin V-FITC and/or PI. (k) The expression of GSDME and GSDME-N was analyzed by western blotting at protein levels. (l) The intensity of bands was quantified by Image J, and β-actin was used as a control.

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