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. 2024 Jun 11;221(8):e20231518. doi: 10.1084/jem.20231518

Figure 4.

Figure 4.

HSV-1 ICP0 is required to inhibit the NLRP1 inflammasome. (A) Kinetics of membrane permeability in primary keratinocytes infected with HSV-1 ∆ICP0 virus 7134 and the corresponding rescue 7134R at MOI 10 followed by treatment with talabostat (30 μM). Data are presented as the mean of three independent experiments ± SEM. Area under the curves was calculated for each treatment and statistical significance was calculated by Student’s t test (*P < 0.05, **P < 0.01). (B) Immunoblots of lysates from primary human keratinocytes infected with HSV-1 ∆ICP0 virus 7134 or the 7134R rescue virus at MOI 10 for 4 h followed by treatment with talabostat (30 μM) for 24 h. Immunoblots were also run from DSS-treated lysates to assess ASC oligomerization and from protein extracted from supernatants to test for caspase-1 cleavage. Images are representative of n = 3 biological replicates. (C) Immunoblots of lysates from primary human keratinocytes infected with HSV-1 ∆ICP0 virus 7134 or the 7134R rescue virus at MOI 10 for 4 h followed by treatment with anisomycin (1 μM) for 24 h. Images are representative of n = 3 biological replicates. (D) Quantification of membrane permeability from control NTERT2G1 keratinocytes (empty vector) or cells expressing dTAG-NLRP1 infected with HSV-1 7134 or 7134R (MOI 10) for 4 h followed by treatment with dTAG-13 or dTAGV-1 for 8 h. Data are presented as the mean of three independent experiments ± SEM. Statistical significance was calculated by Student’s t test (*P < 0.05, ***P < 0.001, ****P < 0.0001). (E) Kinetics of membrane permeability in primary human keratinocytes infected with non-replicative viruses HSV-1 d106 and d109 (MOI 10) for 4 h followed by treatment with talabostat (30 μM). Data are presented as the mean of three independent experiments ± SEM. Area under the curve was calculated for each treatment and statistical significance was calculated by Student’s t test (**P < 0.01, ***P < 0.001, ****P < 0.0001). (F) Immunoblots of lysates from primary human keratinocytes infected with HSV-1 d106 or d109 at MOI 10 for 4 h followed by treatment with anisomycin (1 μM) for 24 h. Images are representative of n = 3 biological replicates. (G) Quantification of membrane permeability from control NTERT2G1 keratinocytes (empty vector) or cells expressing dTAG-NLRP1 infected with HSV-1 d106 or d109 for 4 h followed by treatment with dTAG-13 or dTAGV-1 for 8 h. Data are presented as the mean of four independent experiments ± SEM. Statistical significance was calculated by Student’s t test (***P < 0.001, ****P < 0.0001) Source data are available for this figure: SourceData F4.