Reovirus Activates a Caspase-Independent Cell Death Pathway

Berger, Angela K.; Danthi, Pranav

Antibodies and reagents. Antisera raised against T3D, T1L, and NS have been described (82, 83) . Rabbit antisera specific for cleaved caspase-3 and HMGB1 were purchased from Cell Signaling, rabbit antisera specific for RelA and IB␣ were purchased from Santa Cruz Biotechnology, and rabbit antiserum against poly(ADP-ribose) polymerase (PARP) was purchased from Roche. Goat antiserum specific for Bid was purchased from R&D Systems. Mouse antisera specific for tubulin and PSTAIR were purchased from Sigma. Alexa Fluor-conjugated anti-mouse IgG, anti-rabbit IgG, and anti-goat IgG secondary antibodies were purchased from Invitrogen. IRDye-conjugated anti-guinea pig IgG was purchased from LI-COR. TNF-␣ was purchased from Sigma and used at a concentration of 10 ng/ml. Inhibitors of caspase-8 and -9 (Z-IETD-FMK and Z-LEHD-FMK), purchased from R&D Systems, and a broad-spectrum caspase inhibitor [Z-VAD(OMe)-FMK], purchased from Santa Cruz Biotechnologies, were used at a concentration of 25 M. The RIP1 inhibitor necrostatin-1 was purchased from Santa Cruz Biotechnologies and used at a concentration of 50 M. IKK inhibitor, described by Bayer (BAY- (37) , was used at a concentration of 5 M. The IDO inhibitor 1-methyl-L-tryptophan (1-MT) was purchased from Sigma and used at 10 and 100 M. As shown in the respective experiments, none of the inhibitors displayed any cytotoxicity at the concentration used.