Citations

3.2.3.2 Tract tracing surgeries

Animals were administered a subcutaneous injection of the analgesic butorphanol (2
mg/kg; Wyeth, Madison, NJ, USA) and an intraperitoneal injection of the anesthetic sodium
pentobarbital (90 mg/kg; CEVA Santé Animale, France), their head was shaved, and they were
placed in the stereotaxic frame. The surgical procedure mostly followed that of a previously
described protocol (Yamakawa et al., 2016). The incisor bar was set at 2.0 mm below the
interaural level. Following incision of the scalp, a burr hole was drilled into the skull and a
borosilicate capillary glass tube (with a tip of approximately 28-30 µm tip; with filament,
original outer diameter:1.50 mm, inner diameter: 0.86 mm, Warner instruments, Hamden, CT,
USA) was backfilled with CTβ (1% dissolved in dH2O; List Biological Laboratories Inc,
Campbell, CA, USA). …

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

… Confluent cells were stimulated with LPS from Escherichia coli serotype 0111: B4 (lot no. 2014641; List Biological Laboratories, Campbell, CA) at 100 ng/ml …

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

… BDCA2-DTR mice. After 6 weeks for reconstitution, mice were then depleted of pDCs by intraperitoneal injection of Diphtheria toxin (DT- List Biological Laboratories) every 48 hours for up to 2 weeks …

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

… Sham mice received an intragastric gavage of a bicarbonate buffer (pH 9.0) vehicle containing 10 µg cholera toxin (List Biologicals). …

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

… the MOG solution as well as a 100 µl dose of 2 ng/µl pertussis toxins and virulence factors (List Biological Laboratories Inc., Campbell, CA, USA) diluted in sterile PBS (Thermo Fisher Scientific). Pertussis toxins and virulence factors were administered again on day 2. Pertussis toxins allow an increase in the blood-brain barrier …

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)

… Pertussis Toxin (500ng per mouse; List Biological Laboratories, Inc.) was injected intraperitoneally on days 0 and 2 (6). The animals were monitored daily for EAE signs based on a standard EAE 0 to 5 scale scoring system as follows: …

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)

3.3.3) Experimental autoimmune encephalomyelitis (EAE) induction

Young (8-week) and middle-aged (52-week) female C56BL6 mice (Jackson Laboratories) were subcutaneously injected with 50µg per 100 µL MOG 35-55 peptide (Protein and Nucleic Acid Facility, Stanford University School of Medicine) in complete Freund’s adjuvant supplemented with
4mgmL-1 heat-inactivated Mycobacterium tuberculosis H36a (Sigma-Aldrich). A total of 50 µL emulsion was deposited on either side of the tail base. Pertussis toxin (300 ng per 200 µL; 180, List Biological Laboratories) was intraperitoneally injected on days 0 and 2 after MOG immunization. …

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

Chemicals and reagents

Toxins used for Drosophila feeding assays (Table 1) were purchased from List Biological Laboratories (Campbell, California).
Toxins (product numbers) used for screening in Drosophila included:
Bordetella pertussis toxin (180) and adenylate cyclase toxin (188L), Clostridium difficile toxins A (152C) and B (155B), Clostridium perfringens ε-toxin (126A), Clostridium septicum α-toxin (116L), Corynebacterium
diphtheriae toxin (150), Escherichia coli O111:B4 lipopolysaccharide (201), Pasteurella multocida toxin (156), Salmonella typhimurium lipopolysaccharide (225), Salmonella minnesota R595 lipopolysaccharide (304) and monophosphoryl lipid A (401), and Vibrio cholerae cholera toxin
(100B).

• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #188L – Adenylate Cyclase Toxin, Recombinant from B. pertussis
• Product #152C – Toxin A from Clostridium difficile
• Product #155 – Toxin B from Clostridium difficile
• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae
• Product #201 – LPS from Escherichia coli O111:B4
• Product #156 – Pasteurella multocida Toxin
• Product #225 – LPS from Salmonella typhimurium
• Product #304 – LPS from Salmonella minnesota R595 (Re)
• Product #401 – LIPID A monophosphoryl from Salmonella minnesota R595
• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

EAE Induction

Female C57BL/6J mice at 6-8 weeks old were purchased from Experimental Animal Center of Air Force Military Medical University. EAE was induced with 200 ug MOG35-55 peptide (NJPeptide, China) emulsified in incomplete Freund’s adjuvant (Sigma – Aldrich, USA) added with 5 mg/mL Mycobacterium Tuberculosis H37RA (Difco, USA). Each mouse was immunized with the emulsion subcutaneously into four sites at the back followed by intraperitoneal injection of 200 ng pertussis toxin (List Biological Laboratory). The pertussis toxin was given again at day 2 post-immunization (p.i.). …

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)

Active EAE was induced in 9-12 week-old male and female mice by subcutaneous injection of 150µg of mouse myelin oligodendrocyte glycoprotein peptide (residues 35-55; MOG35-55) (Genemed Synthesis, Inc., San Francisco, USA) emulsified in Complete Freund’s Adjuvant (CFA) (Thermo Fisher Scientific; DF0639606) containing 5 mg/mL heat-inactivated Mycobacterium tuberculosis (Thermo Fisher Scientific; DF3114338). 250 ng of Bordetella pertussis toxin (List Biological Laboratories, Inc. #181) was administered intraperitoneally on the day of immunization and 48 hours later …

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