Citations

Intramuscular injections

All surgeries were conducted under aseptic conditions on P2 neonates anesthetized by deep hypothermia. For induction, pups were placed in latex sleeves gently buried in crushed ice for 3–5 min and maintenance (up to 15 min) was achieved by placing anesthetized pups on a cold pack (3–4 °C). Following small incisions of the skin to expose the targeted muscles, the viral cocktail (or 0.5% Cholera toxin subunit B (CTB) (List Labs) for labeling of the infrahyoids) was injected via a pneumatic dispense system (Picospritzer) connected to a glass pipette (tip diameter ca. 0.1 mm) mounted on a 3D micromanipulator to guide insertion in the desired muscle. …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt

Methods
… Full-length TcdA and TcdB toxins were purchased from List Biological Laboratories Inc. …

• Product #152C – Toxin A from Clostridium difficile

• Product #155 – Toxin B from Clostridium difficile

Stromal Cell Culture

Mouse lung fibroblasts were isolated from whole lung explants by digestion … For experiments, fibroblasts were used between passages 4 and 6. Fibroblasts were serum-starved with DMEM/F-12 medium containing 1% FBS for 24 hr before stimulation with IFN-β and lipopolysaccharide (LPS). Confluent cells were stimulated with LPS from Escherichia coli serotype 0111: B4 (lot no. 2014641; List Biological Laboratories, Campbell, CA) at 100 ng/ml and IFN-β (PBL Interferon Source; Piscataway, NJ) at 100 units/ml. After 4 hr of stimulation, the medium was removed, cells were washed with PBS, and RNA was extracted using RNAeasy Plus Mini Kit (Qiagen).

• Product #201 – LPS from Escherichia coli O111:B4

… Pertussis toxin was purchased from List Biological Laboratories, INC. (Campbell, CA, USA) …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

2.4.1. Retrograde labeling

… A pulled glass pipette (~20 μm outer tip diameter) was attached to the arm of the stereotaxic apparatus. A solution of 0.25% CTB (List Biological Labs, Campbell, CA, USA) in sodium phosphate buffer (pH 7.5) was backfilled through the pipette tip using negative pressure, then a wire connected to a current source (Stoelting) was inserted into the tracer solution. …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt

… For the induction of conventional EAE, naïve female C57BL/6 mice, 8-12 weeks old, were injected subcutaneously (s.c.) in their flanks with 0.1ml of an emulsion containing 100µg of MOG35-55 and 400µg heat-killed M. tuberculosis (H37Ra, Difco) in Incomplete Freunds adjuvant (IFA, Difco) and, the same day and 2 days later, received intravenously (i.v.) 200ng of B. pertussis Toxin (PTX) (List Laboratories) …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

Induction and assessment of EAE

To induce EAE, 8-week-old female C57BL/6 or OptnΔCD11c and DKO mice with their littermates were immunized subcutaneously with 400 μg MOG35–55 peptide (Sangon) emulsified in CFA (Sigma) containing 4 mg/mL heat-killed Mycobacterium tuberculosis H37Ra (BD Biosciences) at day 0. In addition, 200 ng pertussis toxin (List Biological Laboratories) in 0.1 mL PBS was administered intravenously at day 0 and day 1 post immunization per mice.

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

Anthrax lethal factor (LF) and anthrax protective antigen (PA) recombinant nicked from Bacillus anthracis were purchased from List Biological Laboratories, Campbell, CA, USA

Cell cultures and stimulation

Peritoneal macrophages were isolated from mice after being injected intraperitoneally with 3% thioglycollate solution for 4 days. For NLRC4 inflammasome activation, peritoneal macrophages were treated with LFn-Fla or LFn-Fla 3A with PA for the indicated times. For NLRP1b inflammasome activation, the cells (J774, RAW264.7, or RAW-asc) were cultured with LF (2 μg/ml; no. 172C, List Biological Labs) together with PA (2 μg/ml; no.171E, List Biological Labs) for the indicated times. …

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

… pertussis toxin (List Labs, Campbell, CA, USA), …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)