Figure 7.

Formation of single ASC specks in HSV⁽⁺⁾ and HSV⁽⁻⁾ infected human corneal epithelial cells infected with virulent and less-virulent strains of HSV-1. Human primary corneal epithelial cells (hTCEpi cell line) were infected in vitro with HSV-1 laboratory strains McKrae, 17, F, RE or KOS or with clinical isolates KOS79 and KOS63 at MOI of 1 (A,B) or MOI of 0.1 (C,D). Twenty-four hours post-infection, cells were harvested and stained for FAM-FLICA and analyzed by FACS to determine the levels of expression of single and dense ASC specks, a prerequisite for to Caspase-1 activation. (A) Contour plots showing percentage of FAM-FLICA⁺ (activated Caspase-1⁺ cells) expressed in bulk cells. (B) Contour plots showing percentage of infected cells (HSV-1⁺ cells) at (MOI = 1). (C) Contour plots showing the ASC aggregation (comparing ASC-height vs. ASC-area) in HSV-1⁽⁺⁾ cells (upper panels) or in uninfected bystander cells (HSV-1⁽⁻⁾ (lower panels). (D) From the same culture: contour plots showing percentage of infected cells (HSV-1⁺ cells) at (MOI = 0.1). The gating strategy is described in Supplemental Figure S1B. Results are representative of three experiments.