Citations

… goat anti-CTb, List Biological Laboratories, 1:50,000 dilution), in PBS with 0.25% Triton-X and 0.05% sodium azide. Tissue was then washed three times in PBS (30 s each) and incubated in biotinylated secondary (1:1000 dilution, …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

EAE induction:

Mice were immunized subcutaneously with 100 µL of emulsified complete Freunds adjuvant (MilliporeSigma) supplemented with 10 µg/L Mycobacterium tuberculosis H37Ra (BD) and 100µg MOG₃₅₅₅ peptide (Charit Berlin) and received intraperitoneal injections of 200 ng pertussis toxin (List Biological Laboratories) at the time of immunization and 48 hours later. …

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)

… In addition, the animals received intraperitoneal (ip) injection of 150 ng of pertussis toxin (PT; List Biological Laboratories, RRID: SCR_014324) on days 1 and 2. …

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)

Mouse model of peanut-induced anaphylaxis

… Mice were sensitized intragastrically with 1.8 mg of PE mixed with 15 μg of cholera toxin (List Biological Laboratories, Campbell, Calif) in 300 μL of PBS on days 0, 1, 2, 7, 14, 21, and 29 (7 groups, 4-6 per group). …

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

… 1mg/ml pertussis toxin (PTX; List Biological Laboratories, 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)

… Mice were immunized subcutaneously with 200µg of MOG3555 in complete Freunds adjuvant (Sigma-Aldrich) containing 500µg of killed M. tuberculosis strain H37Ra (Difco) and received an intraperitoneal injection of 200ng of pertussis toxin (List Biological Laboratories) at the time of immunization and 48h later. …

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)

… For intoxication of cells, media was replaced over 105 cells previously seeded in a 12-well plate. 31.7 nM PA alone (List Labs, #171E) or …

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

2.3 Immunizations

2.3.1 Intragastric immunizations

Groups of mice were immunized intragastrically on two consecutive days at bi‐weekly intervals with OD 0.5 of formalin‐killed whole cell bacteria corresponding to 1 × 109 bacteria/mL and 7.5 μg of CT (List Biological Laboratories) or 15 μg of mmCT12 with bicarbonate buffer on each occasion (hereafter referred to as IG + CT or IG + mmCT) (Figure 3A).

2.3.2 Sublingual immunizations

Groups of mice were immunized sublingually twice at a bi‐weekly interval with 200 μg of H pylori lysate antigens reconstituted in 7.5 μg of CT or 15 μg of mmCT on each occasion (hereafter referred to as SL + CT or SL + mmCT) (Figure 3A). H pylori vaccination regimen (route and vaccine) was carefully chosen based on our previous studies.14, 18

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

Immunizations

To induce local IgA immunity mice were orally immunized with 10 mg of OVA protein grade V (Sigma) dissolved in 0.5 mL of PBS and containing 3% (w/v) NaHCO3 with or without the addition of 10 µg cholera toxin (CT) adjuvant (List Biological Laboratories, Campell, CA) to each dose. The mice were given three oral administrations, 10 days apart, using intragastric intubation. Parenteral immunizations in 0.5 mL PBS were done i.p twice with 50 µg OVA w/wo 2 µg CT.

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

Wt-RABV tracing:

For the present trans-synaptic tracing (Figure 1), nine out of eleven cases injected were used from the previous study, in which we focused on cerebellar connections into sensorimotor cortices (Aoki et al., 2019). Before surgery, animals were anesthetized with ketamine (80 mg/kg, Imalgene, France) and xylazine (10 mg/kg, Bayer, Germany). Appropriate levels of anesthesia were monitored by the absence of whisker movements and foot-pinch withdrawal reflex. Additional doses of the ketamine-xylazine mixture were administered i.p. when necessary. After placing animals into a stereotaxic frame (David Kopf Instruments), a mixture (0.15 or 0.2 L) of the French subtype of CVS-11 rabies virus (Wt-RABV, Aoki et al., 2019; Coulon et al., 2011; Raux et al., 1997; Ruigrok et al., 2008; Suzuki et al., 2012; Ugolini, 2010) and cholera-toxin b subunit (CTb, low salt; List Biological Laboratories, 1% w/v in 0.2 M phosphate buffer (PB) at pH 7.4: the injection solution consisted of four parts Wt-RABV and one part CTb) was injected by a 1 l Hamilton syringe in the following functional areas of the cerebral cortex: primary motor cortex (M1, n = 4), secondary motor cortex (M2, n = 4), and medial prefrontal cortex (mPFC, n = 3, one centered on prelimbic and two centered on cingulate cortex). As shown in our previous study, coordinates of the injection sites were determined by reference to the standard rat brain atlas (Aoki et al., 2019; Paxinos and Watson, 2004). The viral stock was kept at 80C until use. This CVS-11 strain of RABV has been confirmed to be transported trans-synaptically in a retrograde direction as well as in the time-dependent manner (Aoki et al., 2019; Kelly and Strick, 2004; Ruigrok et al., 2008; Suzuki et al., 2012; Ugolini, 2010). No neighboring neurons are infected unless they have synaptic contacts to the already infected cells. Adding CTb to the injection solution enabled accurate determination of the injection site (Aoki et al., 2019; Suzuki et al., 2012). Upon the injection, the needle was left in place for another 5 min to allow the virus to spread. After surgery, animals were monitored for signs of stress or discomfort. …

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