Citations

… DT powder is purchased from Sigma-Aldrich #D0564 or List Biological Labs #150 and can be stored at 2-8 °C …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

Cell culture and treatment with S100A4

BMDCs were generated by culturing bone marrow cells in culture medium (RPMI-1640; Sigma-Aldrich) supplemented with 10% fetal bovine serum (Gibco), 1% L-glutamine, 1% gentamicin, and 50 μM 2-mercaptoethanol (all from Sigma-Aldrich) in the presence of 200 ng/ml recombinant Flt3L (PeproTech) for nine days. Cells were incubated in vitro with various stimuli, including CT (List Biological Laboratories) (0.1 μg/ml), …

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Stereotaxic injections and retrograde tracing

Mice used in the experiments were Skor2GFP/wt (n=6). Mice were anesthetized with isoflurane, attached to the stereotaxic frame, and a small hole was drilled into their skull. For retrograde tracing of vlPAG neurons, bilateral, intracranial injections of 0, 2% Cholera toxin B subunit (#104; List Biological Lab.Inc.) were injected at the speed of 50 nl/min using a microinjector (UltraMicroPump III, World Precision Instruments) and microsyringe (Hamilton 7803-06). The stereotaxic coordinates for injections were (measured from bregma, in mm): -5,19 to -5,4 (AP); 0,88 (ML); -4,4 (DV). The coordinates were obtained from the mouse brain atlas (Paxinos and Franklin, 2012). Mice were intracardially perfused 5-7 days after the injections and the brains were collected. Brains were processed for vibratome sectioning and further for IHC stainings and analysis for GFP and CtB expression in the midbrain.

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

Flow cytometry

… Cryopreserved liver or PBMCs were thawed and rested overnight in sterile R10 media (RPMI 1640, Corning), supplemented with 10% fetal bovine serum (Gemini Bio-Products, West Sacramento, CA), penicillin/streptomycin, and L-glutamine plus 10 U/mL DNAse I (Roche Life Sciences, Indianapolis, IN) at 37°C, 5% CO2, and 95% humidity incubation conditions. Cells were stimulated as previously described,45 with a final concentration of 2 mg/mL AAV8 peptide pool C, hOTC peptide pool A or 1 μg/mL Staphylococcal enterotoxin B (SEB) (List Biological Laboratories, Campbell, CA) as a positive control. …

… Incomplete Freund (DF0639-60-6) and Mycobacterium tuberculosis (DF3114-33-8) from Difco (Detroit, MI, USA). Pertussis toxin was obtained from List Biological Lab. (#181)…

• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)

STAR★Methods
Key resources table

Diphtheria toxin (mutant CRM197) List Biological Laboratories Cat. no. 149A

Enzyme linked immunosorbent assay (ELISA)

An ELISA to determine the development of antibodies against diphtheria toxin (DT) after long time DT treatment was performed according to Buch et al. (Buch et al., 2005). For the assay, a 96-well plate was coated with 2μg/mL DT (mutant CRM197, List Biological Laboratories) at 4°C overnight and the free binding sites of the plate were blocked the following day with 0.5% BSA in PBS for two hours at RT. …

• Product #149A – Diphtheria Toxin CRM197, Mutant

… LPS was obtained from List Biological Laboratories…

Author did not specify which List Labs LPS product was utilized in their research.
List Labs provides the following LPS products: https://listlabs.com/product-information/lipopolysaccharides/

… (1% CTB, 10 µl; List Biologicals) unilaterally into the ECR using a Hamilton syringe using a 7-d survival time, similar to our previous study (45). CTB labeling was visualized using a primary antibody to CTB (goat anti-CTB; 1:1,000; List Biologicals), …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt
• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

… Binding of radiolabeled LT-IIc B-subunits, and cholera toxin B-subunits (CTB) (List Labs., Campbell, CA), to glycosphingolipids on thin-layer chromatograms was done as described previously (Ångström et al. 2000)…

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

Induction of EAE

To induce active EAE, 9–10-week-old female C57BL/6J mice were immunized subcutaneously with 200 µg MOG35–55 peptide (MBL, Tokyo, Japan) and 400 µg Mycobacterium tuberculosis in incomplete Freund’s adjuvant (Difco Laboratories). Pertussis toxin (200 ng per mouse; List Biologicals) was injected intraperitoneally the same day and 2 days 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)