Citations

EAE induction and orexin A treatment:

EAE was induced by subcutaneous immunization in the flanks with 100µg of MOG₃₅₅₅ (GL Biochem, China) emulsified with complete Freunds adjuvant (CFA) supplemented with 5 mg/mL of Mycobacterium tuberculosis H37Ra (Difco, USA). In addition, mice received intraperitoneally (IP) 300 ng of pertussis toxin (List Laboratories, USA) on days 0 and 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)

…Under anesthesia with isoflurane, a total of 4 µL of 1% CTB (List Biological Labs, Campbell, CA) solution was injected slowly into LI 4 or LR 3 on the left side. The depth of the injection was 2-3 mm. FIG. 1. LI 4 (A) and LR 3 (B) in rat forefoot and hind foot respectively…

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

Briefly, we isolated a subset of birds in experimental cages the day before LPS injection, then dosed them the following morning with an intra-abdominal injection of 1 mg ml1 LPS from E. coli (O55:B5; List Biological Laboratories, Campbell, CA, USA) dissolved in PBS. …

• Product #203A – LPS from Escherichia coli O55:B5

Infections and stimulations:

DCs were infected or stimulated with innate agonists between day 4 and day 6 after differentiation. DCs were counted on day 4 and resuspended at 800,000 cell per ml in fresh medium with GM-CSF, IL-4, and polybrene (1 g/ml) and then reseeded into appropriate culture vessels. For most assays, DCs were infected at a density of 800,000 cells per ml by diluting virus in DC media (without cytokines or polybrene) to a final volume normalized to controls. Innate and inflammatory stimuli were used at the indicated concentrations: polyI:C (InvivoGen, 10 g/ml); LPS (List Biological Laboratories, INC, 1 ng/ml).

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/

Chemicals and Reagents:

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

EAE in Mice:

Female C57BL/6 mice at 1012 weeks of age were immunized with subcutaneous injection of 200µg MOG35-55 peptide and 400-g mycobacterium tuberculosis H37Ra in a 200µL emulsion. Te mice received intraperitoneal injection of 500-ng pertussis toxin on Day 0 and Day 2. On Day 7 afer immunization or the onset of EAE symptoms, the mice were divided into groups and treated for 14 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)

… agents were added 60 min before the challenge with lipopolysaccharides (LPS) (10 ng/ml, Escherichia coli 055:B5; List Biological Laboratories …

• Product #203A – LPS from Escherichia coli O55:B5

Transganglionic tracer injection:

DCN labeling using transganglionic axonal tracers was performed as described in our previous report⁹. Briefly, rats (n=3) were anesthetized with intraperitoneal injections of Ketamine (75mg/kg, Bioniche, Morgantown, WV)/Dexmedetomidine (0.25mg/kg, Dexdomitor, Pfizer, New York, NY) and DCNs at T7 and T13 spinal levels were exposed through a skin incision along the median line across thoracic areas. Transganglionic axonal tracers, IB4 (Vector Laboratories, Burlingame, CA) and CTB (List Biological Laboratories, Campbell, CA) were dissolved in sterile phosphate buffered saline (PBS; pH 7.4) for injections (1 ul of 2% solution for each tracer). IB4 was injected into left DCNs and CTB into right DCNs, at both T7 and T13 spinal levels. A 30-gauge needle with a Hamilton syringe (75RN, Hamilton, Reno, NV) was inserted into a proximal site of each DCN, and injections were made using a motorized pump (Model #310, Stoelting Co., Wood Dale, IL) for over 1minute. Then, the skin was closed with surgical staples. Animals were recovered from anesthesia with a reversal drug, Antisedan (2mg/kg, Pfizer), and they received post-operative care including saline injections as needed with daily monitoring.

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

Immunohistochemistry:

Free-floating sections were immunostained for CTb, NeuN or cFos by overnight incubation in goat anti-CTb (List Biological Laboratories, 7032A9, 1: 50,000 dilution), mouse anti-NeuN (Millipore, MAB-377, 1: 5000 dilution), or rabbit anti-cFos (Millipore, ABE457, 1:1000 dilution) primary in PBS with 0.25% Triton-X and 0.05% sodium azide. Afterwards, tissue was washed three times in PBS and incubated in biotinylated donkey-anti-goat, anti-mouse or anti-rabbit secondary (1:1000 dilution, Jackson Immunoresearch, West Grove, PA) for 30 min, followed by three 30 s rinses in PBS, followed by 1 hr in avidin-biotin complex (Vector). For TH-staining, tissue was then rinsed in sodium acetate buffer (0.1M, pH 7.4), followed by incubation for 5 min in 1% diaminobenzidine (DAB). For cFos and NeuN staining, nickel and hydrogen peroxide (Vector) were added to reveal a blue-black reaction product.

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

… Goat anti-Cholera Toxin B List Biological List Biological Cat# 703; …

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

Efferocytosis Assay:

Efferocytosis was assessed by flow cytometry [68]. Briefly, 5 10⁵/well monocyte-derived human M2(IL-4+IL-10+Dex) macrophages or M2(Dex) macrophages were preincubated with 10 nM recombinant PA (List Biological Laboratories, Campbell, CA, USA), 10 nM recombinant EF (List Biological Laboratories), or various concentrations of PA + EF (ET) for 4 h at 37 C in 5% CO₂ atmosphere. Fluorescently-labeled apoptotic human PMN were preincubated with 10% serum, Protein S, or MFGE8 for 1 h unless otherwise noted and then added to the macrophages, resulting in a macrophage:PMN ratio of 1:5. After 1 h at 37 C in 5% CO₂ atmosphere, the percentage macrophages containing intracellular (eFluor 670⁺) but not surface-bound (surface CD66b) PMN was assessed by flow cytometry.

• 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).