Bystander Killing during Avian Leukosis Virus Subgroup B Infection Requires TVB ^S3 Signaling

Anthrax Lethal Toxin Kills Macrophages in a Strain-Specific Manner by Apoptosis or Caspase-1-Mediated Necrosis

“…TUNEL assays were performed as previously described. 39 For analysis of membrane integrity, cells were detached in a solution of 2.6 mM TO-PRO-3 (Invitrogen, Carlsbad, CA) in PBS. TO-PRO-3 was quantified by flow cytometry.…”

Mitochondrial Impairment Mediates Cytolysis in Anthrax Lethal Toxin-Treated Murine Macrophages

“…Fifty µg of protein from each sample was loaded on precast Biorad SDS-Tris HCl polyacrylamide gels, and transferred to PVDF membranes (Amersham) as described previously. 28 Membranes were probed with the following antibodies: anti-ubiquitin polyclonal antibody (Sc-9133; Santa Cruz Biotechnology), anti-MEK-3 polyclonal antibody (Sc-960; Santa Cruz Biotechnology), anti-p35 polyclonal antibody (Sc-820; Santa Cruz Biotechnology), anti-SDH polyclonal antibody (Sc-25851; Santa Cruz Biotechnology) and anti-actin monoclonal antibody (Ac-40; Sigma). We used a polyclonal hrp-conjugated anti-rabbit antibody as a secondary antibody (Sc-2313; Santa Cruz Biotechnology), except for an hrp-conjugated anti-mouse antibody in the case of actin (NXA931; Amersham).…”

“…Coomassie Blue staining (Invitrogen) and Western blotting was performed as described previously. 28 In brief, cells were lysed in the wells with caspase-3 lysis buffer (R&D Systems) supplemented with protease cocktail inhibitors (Roche) and 1 mM PMSF (Sigma). Cellular lysates were centrifuged, and supernatants were mixed with SDS sample buffer.…”

Proteasome Inhibitors Prevent Caspase-1-Mediated Disease in Rodents Challenged with Anthrax Lethal Toxin

“…15 Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and propidium iodide exclusion assays were performed as previously described. 25,26 For analysis of caspase-1 activation, we used a fluorescent caspase-1 activity assay, FLICA, from Immunochemistry Technologies (Bloomington, MN). The assay was performed in 96-well plates, with 8 ϫ 10 4 cells/well.…”