FIG 2.

Reovirus can induce necroptosis in primary BMDMs. (A) BMDMs from wild-type mice were mock infected or infected with 50 PFU/cell of T3D in the presence of DMSO, Z-VAD-FMK (25 μM), Nec1 (50 μM), or both inhibitors. Cell death 48 h following infection was assessed by using the Cell Titer Glo system. Luminescence measurement in similarly treated, uninfected cells was considered to represent 100% viability. *, P < 0.05 compared to DMSO-treated cells. (B) BMDMs from wild-type (left) or RIP3^−/− (right) mice were mock infected or infected with 50 PFU/cell of T3D in the presence of DMSO or Z-VAD-FMK (25 μM). Cell death 48 h following infection was assessed by using the Cell Titer Glo system. Luminescence measurement in uninfected cells of the same genotype that were similarly treated was considered to represent 100% viability. *, P < 0.05 compared to DMSO-treated cells of the same genotype. (C) BMDMs were infected with 50 PFU/cell of T3D in the presence of DMSO or Z-VAD-FMK (25 μM). Cell viability was assessed by Sytox green staining. *, P < 0.05 compared to DMSO-treated cells of the same genotype. (D) BMDMs from wild-type, RIP3^−/−, or Casp8^−/− × RIP3^−/− mice were infected with 50 PFU/cell of T3D. Cell death 48 h following infection was assessed by using the Cell Titer Glo system. Luminescence measurement in mock-infected cells of the same genotype was considered to represent 100% viability. *, P < 0.05 compared to wild-type cells. (E) BMDMs from wild-type or RIP3^−/− mice were infected with 50 PFU/cell of T3D in the presence or absence of Z-VAD-FMK (25 μM). The virus yield 24 h following infection was measured by using a plaque assay. WT, wild type.