Citations

Animals:

Animals were maintained in a pathogen-free environment on a 12-hour light/12-hour dark cycle with full access to food and water. SftpcCreERT2mice (59), obtained from Harold Chapman (UCSF, San Francisco, California, USA), were crossed to Rosa26-mTmG (abbreviated mTmG) mice (The Jackson Laboratory). Male SftpcCreERT2^+/;mTmG^+/ mice were given 20 mg/ml tamoxifen in corn oil at a dose of 0.25 mg/g body weight i.p. every other day for 3 doses at age 6 weeks. Recombination efficiency is shown in Supplemental Figure 1. At 12 weeks of age, mice were treated with 40 g LPS (E. coli 0111:B4, List Biological Laboratories) in 50 l 0.9% saline by intratracheal instillation or left untreated. Mice were randomly assigned to treatment groups. Mice were euthanized 7 or 27 days later. Sprague-Dawley rats aged 24 months were purchased from Charles River or obtained from the University of Michigan Rodent Recycling Program. Tgfbr2^fl/fl mice were purchased from The Jackson Laboratory.

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

EAE:

… In addition, the mice received 400 ng pertussis toxin (List Biological Laboratories) intraperitoneally (i.p.) on days 0 and 2 after immunization. …

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)

EAE Induction and behavioural score:

Mice were given a flank subcutaneous injection of 150 µg of MOG_35-55 emulsified with complete Freunds adjuvant (MOG/CFA) which contained 0.3 mg of heat-inactivated Mycobacterium tuberculosis (H37RA; Difco Laboratories). Each animal also received pertussis toxin (200ng i.p; List Biological Laboratories) on day 0 and day 2 post-immunization. …

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)

EAE induction and oral GlcNAc/GlcNAc-6-Acetate treatment:

… Injections were distributed over two spots on the hind flank. All mice received 400 ng of pertussis toxin (List Biological Laboratories) by intra-peritoneal injection on days 0 and 2 after immunization. …

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)

EAE:

… Pertussis toxin e.g. List biological laboratories, Inc. 180 …

• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer

EAE induction and scoring:

6 to 8 weeks old, sex-matched NLK^fl/fl and NLK^Treg mice littermates were injected subcutaneously in the rear flank with 100 ug MOG^35-55 peptide (2HNMEVGWYRSPFSRVVHLYRNGK-COOH) in complete Freunds adjuvant (CFA) (Sigma) on day 0, and 250ng pertussis toxin (List Biological) was injected intraperitoneally on day 0 and day 2 post-induction. Mice were monitored and disease severity was scored every two days. …

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)

… Recombinant EF was produced as previously described.  Activated recombinant PA (PA63) was from List Biological Laboratories (Campbell, CA). …

• Product #174 – Anthrax Protective Antigen, Activated (PA 63) from B. anthracis

Experimental procedures –

Materials:

Recombinant anthrax toxins PA83, PA63, LF and EF were from List Biological Laboratories (Campbell, CA) …

• Product #174 – Anthrax Protective Antigen, Activated (PA 63) from B. anthracis

Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169L (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

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

Author did not specify which List Labs Anthrax Edema Factor was utilized.  List Labs provides Product #178A(Anthrax Edema Factor (EF), Recombinant from B. anthracis) and Product #173B (Anthrax Edema Factor (EF), Recombinant, Mutant S447N from B. anthracis).

… Then, the mixture was injected subcutaneously into neck (0.1 mL) and both hindlegs (0.05 mL each) of mice. Pertussis toxin (200 ng; List Biological Lab, Inc) was administered intravenously on days 0 and 1 post-immunization. …

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)

C. difficile cytotoxin assay:

A cytotoxicity assay was performed as previously described with the following modifications (25). Briefly, ATCC CCL-81 Vero cells were grown to confluence in Dulbecco modified Eagle medium (Gibco catalog number 11965) supplemented with 10% fetal bovine serum (catalog number 16140) and 1% penicillin-streptomycin (Gibco catalog number 15070). Cells were plated to a density of 10 5 cells/well. Mouse cecal content was diluted 1:10 in sterile PBS, passed through a 0.22- m filter, and serially diluted to 10:6. Filtered samples were tested in duplicate with a corresponding control to which both antitoxin (Techlab catalog number T5000) and sample were added. A positive control of C. difficile TcdA (List Biologicals catalog number 152C) was used. Samples were incubated overnight at 37C, and the cytotoxic titer was determined as the reciprocal of the highest dilution that produced 80% cell rounding.

• Product #152C – Toxin A from Clostridium difficile