Citations

… Anthrax lethal toxin List Biological Laboratories Inc. #171B/#172B …

Treatments

See Table S1 for list of Figures/panels in each figure and corresponding dose and exposure time. General information for the methods is provided here. To induce cytokine release from macrophages, microglia and BMDM cells were primed for 4–5 h (or time indicated in Table S1)with 1 μg/mL of LPS in culture medium, and some cells were pre-treated with 1 μM of CP456773/MCC950 for 30 min before ATP stimulation. Cells were then pulsed with 5 mM ATP for 30 min. For cytokine induction by anthrax lethal toxin (LeTx), LPS-primed cells were treated with LeTx at a final concentration of 250 ng/mL (LF and PA) for 3 h. Cell supernatants were collected and frozen at −80°C until assayed.

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis

… goat anti-EF (773L, List Labs, Campbell, CA, USA), …

… For the activation of NLRP3, nigericin was used at a final concentration of 5 µM. PA (List Biological Laboratories) and LFn-PrgJ (expressed and isolated at Nodthera) were added to yield final concentrations of 1 and 4 …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Animal challenge

Mice were randomly assigned to the control or experimental groups. Mouse TNF (CF09, Novoprotein, China)
and/or LT (LF (172C, List Biological Labs, USA) and PA (171E, List Biological Labs)) diluted in 200 μL of phosphate buffered saline (PBS) was intravenously injected into the mouse tail-vein with or without inhibitors of caspase (Z-VAD-FMK, 627610, Calbiochem, Germany) or p38α (TAK-715, HY-10456, MedChemExpress, USA) diluted at indicated doses. Mouse survival was monitored and recorded at indicated time points.

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Inflammasome activation

Late-log cultures of Salmonella enterica serovar Typhimurium strain SL1344 (a kind gift from Dr. Brad T. Cookson, University of Washington) were grown in L-broth containing 0.3 M NaCl with shaking at 37 °C. Bacteria were washed, resuspended in PBS, and added at a multiplicity of infection of 10 bacteria per macrophage. For activation of NLRP1b, BMM from BALB/c mice were exposed to 1 µg/mL anthrax lethal toxin, prepared as a 1:1 mix of protective antigen and lethal factor (List Biological). …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

… Anthrax Protective Antigen List Biological Laboratories Cat# 171E …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

4.1. Cell Line and Reagents for Development of the AP-1 Reporter Assay
Murine hepatoma Hepa1c1c7 cells (CRL-2026™) purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) were cultured in an alpha minimum essential medium (MEM), supplemented with 10% FBS, 100 IU/mL penicillin, 100 µg/mL streptomycin, and 2 mM L-glutamine. Cultures were maintained at 37 °C in an incubator with 5% CO2. Lyophilized recombinant protective antigen (PA) and lethal factor (LF) were purchased from List Biological Laboratories, Inc. (Campbell, CA, USA), and reconstituted in sterile water to make stock solutions with a final concentration of 1 mg/mL for LF and 2 mg/mL for PA. The stock solutions were then diluted 10 times with 50% glycerol in sterile water to make stock solutions with a final concentration of 0.1 mg/mL for LF and 0.2 mg/mL for PA. …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

Culture supernatant extract anthrax vaccine and its liquid and dry powder formulations preparation and characterization

… Total protein concentrations of the CSE vaccine were measured by bicinchoninic acid assay (BCA, Gene-Star, Beijing, China). An enzyme-linked immunosorbent assay (ELISA, Dakewe, Beijing, China) was used to quantify the PA protein in the CSE vaccine according to a previously method [Citation20]. Serially diluted commercial recombinant PA (rPA, 171E, List Labs, Campbell, CA, USA) was the reference standard. …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

2.1.1. Experimental Materials

The following materials were used: anthrax protective antigen (PA), from List Labs in California, USA, stored at 4 °C; anti-protective antigen (Anti-PA) monoclonal antibody, from List Labs in California, USA, stored at 4 °C; …