Citations

For EAE induction, mice were s.c. immunized with 115 µg MOG 3555 peptide (Washington Biotech) emulsified in CFA (Difco) supplemented with 400 µg/mL Mycobacterium tuberculosis (Difco). On days 0 and 2 postimmunization, mice were i.p. injected with 300 ng pertussis toxin (List Biological).

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)

… or 30% CB-FITC (List Biological Laboratories Inc., Campbell, CA) (all diluted in artificial cerebrospinal fluid …

• Product #106 – Cholera Toxin B Subunit (CTB) from Vibrio cholerae FITC, Conjugate

Cell stimulation:

Cells were infected with H1N1 inuenza A/Puerto Rico/8/34 (PR8; Charles RiverLaboratories) (orH3N2inuenzaX31where indicated;PaulThomas, St. Judes, Memphis, TN) at a multiplicity of infection (MOI) of 10 virions/DC and encephalomyocarditis virus at an MOI of 2. Cells were treated with 30 mM poly(I:C) (Invitrogen), 30 mM R848, 1 mg/ml LPS (List Biological), …

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/

Antibody Characterization:

Goat polyclonal antiserum against cholera toxin b subunit (CTb) (List Biological Laboratories, Campbell, CA). Kishi and colleagues (Kishi et al., 2000) demonstrated that this antibody stains CTb injection sites and retrogradely labeled neurons in the rat brain. As well, in present study this antibody clearly stained CTb injection sites and retrogradely labeled neurons in the mouse brain, but it did not stain brain sections from animals without CTb injections (data not shown), indicating that this antibody has no cross-reactivity to any of endogenous antigens in the mouse brain. The antibody is registered in the JCN Antibody Database and has been frequently and successfully used in many other studies (Al-Khater et al., 2008; Howorth et al., 2009; Wittmann et al., 2009)

Stereotaxic surgery for retrograde neuronal tracing:

Surgical procedure was followed as previously described (Chamberlin et al., 1998; Gautron et al., 2010a). Briefly, MC4R-GFP mice were anesthetized with ketamine HCl/xylazine HCl (80:12 mg/kg, i.p.) and restrained in a Kopf (Tujunga, CA) stereotaxic apparatus. A small hole was drilled into the skull under aseptic conditions. A glass micropipette connected to an air pressure injector system or iontophoresis machine was positioned via the stereotaxic manipulator. Approximately 50100 nL of retrograde tracer, fluorogold (Fluorochrome, CO, USA) or cholera toxin subunit B (CTb) (List Biological Laboratories), was slowly administered into the ventral tegmental area (VTA) (3.2 mm from bregma, 1.1 mm lateral with 10 degree injection arm, …

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

… Goat anti-CTB was made by List Biological Labs (Campbell, CA). …

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

Supplemental Experimental Procedures:
 
Cell culture Bone marrow-derived macrophages were generated from femur exudates of mice by culture for 7 days at 37C in 5% CO2 in DMEM (Invitrogen) supplemented with 10% FCS, 5 mM HEPES, 0.2 mg/ml L- glutamine, 0.05 mM 2-ME, 50 mg/ml gentamicin sulfate, and 10,000 U/ml penicillin and streptomycin with 30% L cell-conditioned medium. Macrophages were then washed with PBS containing 1 mM EDTA, suspended in phenol red-free antibiotic-free DMEM supplemented with 5% FCS, activated with 100 ng/ml ultrapure LPS from Salmonella Minnesota (List Biologicals), and allowed to adhere for approximately 18 hr.

• Product #434 – ULTRA PURE LPS from Salmonella minnesota R595 (Re)

EAE:

… Pertussis toxin (200 ng in PBS; List Biological Laboratories) was injected intraperitoneally on days 0 and 2 after immunization. EAE clinical symptoms were scored daily 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)

Surgery:

To trace the input to the MPB, six rats received brain microinjection of 1.0 % Cholera Toxin subunit B (CTB; List Biological) within the MPB [coordinates: anteroposterior (AP) 9.4 mm, lateral (L) 1.7 mm, dorsoventral (DV) 5.8 mm, as per the rat atlas of Paxinos and Watson (Paxinos and Watson, 2005)]. …

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

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

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)

Measurement of Intracellular Calcium:

Cells (HFFs, HASMC, U373MG) were infected with HCMV recombinants at an MOI of 3 for 48 hours, then loaded with Fluo4-AM (Molecular Probes) for 60 min at 37°C. Pertussis toxin (PTx) was used at a concentration of 200 ng/mL to treat infected cells 24 hours prior to loading (List Biological 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)