Inhibitors of STAT3, caspase-1 and RIP1K enhance vaccinia virus infection and preserve the viability of keratinocytes, but not Vero cells.

<p>A) Detection of luciferase activity over 24 hours post infection with ACAM-luc at 0.2 MOI in HEK001 and 308 keratinocytes, and Vero cells. Mean of replicate wells is plotted. B) Infection of keratinocytes (HEK001, 308) or Vero cells was performed with ACAM-2000 at 20 MOI. Viability at 6 hours post-infection was assessed by Alamar blue dye conversion (n = 4). C) Infection of HEK001 was performed with ACAM-2000 at 20 MOI, with vehicle control, Stattic, or caspase-3 inhibitor added. Viability at 6 and 12 hours post-infection was assessed by Alamar blue dye conversion (n = 4). D) Infection of keratinocytes (HEK001, 308) or Vero cells was performed with ACAM-2000 at 20 MOI, in the presence of DMSO vehicle control, STAT3 inhibitor, caspase-1 inhibitor, or Nec-1. Viability at 24 hours was assessed by Alamar blue dye conversion (n = 4). E) Keratinocytes (HEK001, 308) or Vero cells were infected with ACAM-luc at 0.2 MOI, in the presence of DMSO vehicle control, STAT3 inhibitor, Nec-1, or caspase-1 inhibitor. Luciferase activity at 24 hours is expressed as percent of uninfected control (n = 4). F) Differentiated primary keratinocytes at the air-liquid interface were infected with ACAM-luc at 20 MOI, in the presence of DMSO, STAT3 inhibitor, or Nec-1 (n = 2). Representative of triplicate experiments is shown. Asterisk, p≤0.05.