Fig. 2: Characterization and verification of VNP-GD-induced tumor cell pyroptosis. | Nature Communications

Fig. 2: Characterization and verification of VNP-GD-induced tumor cell pyroptosis.

From: Enhancing Gasdermin-induced tumor pyroptosis through preventing ESCRT-dependent cell membrane repair augments antitumor immune response

Fig. 2

a Direct observation of the pyroptosis of 4T1 cells under confocal microscope after different treatments with PBS, VNP, GD (GSDMD protein cage), VNP-GD (GSDMD protein cage-conjugated VNP), and VNP-GD+EI-NP (GSDMD protein cage-conjugated VNP + EI-NP) for 24 h. 4T1 cells were further stained with Annexin V for imaging. Scale bar: 15 µm. The experiments were repeated three times independently. Flow cytometry analysis of cell uptake of SYTOX green (b) and PI (c) in 4T1 tumor cells after incubation with PBS, VNP, GD (GSDMD protein cage), VNP-GD (GSDMD protein cage-conjugated VNP), and VNP-GD+EI-NP (GSDMD protein cage-conjugated VNP + EI-NP) for 24 h. d Data analysis of the PI-positive cells after different treatments. Data are presented as mean ± s.d. (n = 3 biologically independent samples) and analyzed with one-way ANOVA followed by Dunnett’s multiple comparisons test. VNP-GD + EI-NP vs. VNP-GD: **P = 0.0055; VNP-GD + EI-NP vs. GD: ****P < 0.0001. LDH (e) and HMGB1 (f) release after the treatments of PBS, VNP, GD (GSDMD protein cage), VNP-GD (GSDMD protein cage-conjugated VNP), and VNP-GD+EI-NP (GSDMD protein cage-conjugated VNP + EI-NP) for 24 h. Data are presented as mean ± s.d. (n = 3 biologically independent samples) and analyzed with one-way ANOVA followed by Dunnett’s multiple comparisons test. (e VNP-GD+EI-NP vs. VNP-GD: **P = 0.0068; VNP-GD+EI-NP vs. GD: ***P = 0.0001; f VNP-GD+EI-NP vs. VNP-GD: *P = 0.0119; VNP-GD + EI-NP vs. GD: ***P = 0.0007). Source data are provided as a Source Data file.

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