Citations

2.8. EAE and Spinal Cord Histology

All experiments were conducted with ethics approval from the Animal Care Committee at the University of Calgary under guidelines of the Canadian Council of Animal Care. Eight-to-ten-week-old C57BL/6 female mice (Charles River Laboratories, Montreal, QC, Canada) with an average weight of 22 g were immunized subcutaneously with 50 μg/100 μL of myelin oligodendrocyte glycoprotein (MOG) 35–55 peptide (Protein and Nucleic acid facility, Stanford University, USA) in CFA supplemented with 4 mg/mL heat-inactivated Mycobacterium tuberculosis H37Ra (Sigma). This method has been previously described [21,47]. A 50 μL emulsion was deposited on either side of the tail base. Pertussis toxin (PTX) (300 ng/200 μL, List Biological Laboratories, Campbell, CA, USA) was injected intraperitoneally on days 0 and 2 after MOG 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)

Reagents

…Recombinant PA was purchased from List Biological Laboratories, Inc. (Campbell, CA, USA) …

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

… Then all mice were treated twice with 200 ng pertussis toxin (List Biological Laboratories, USA) in total by intraperitoneal injection at 0 and 48 h after 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)

Experimental autoimmune encephalomyelitis (EAE) models

MOG35-55 peptide was used to immunize 12 female C57Bl/6J mice with a dose of 150 μg (Peptide international)mixed with complete Freud’s adjuvant (CFA; SigmaAldrich; USA) in concentration of 10 mg/ml heat-killed Mycobacterium tuberculosis H37Ra (Difco Laboratories). Pertussis toxin were injected into mice intraperitoneally (List Biological Laboratories; 500 ng) at day 0 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)

STAR★Methods
Key resources table

Lipopolysaccharide, e-coli-O11:B4-derived (LPS) List Labs #201

Murine primary cell cultures

Peritoneal cells were isolated from naïve mice by lavage with 10mL of PBS containing 0.5mM EDTA. Lavage was estimated to contain 50% macrophages. Peritoneal macrophages were isolated by adhesion purification of lavage cells in RPMI-10 (RPMI-1640 containing 10% heat-inactivated fetal bovine serum (v/v), 1X Pen/Strep/Glut, 1mM HEPES, and 100μM Sodium Pyruvate) plated 1.3×106 macrophages/well in 6-well plates for FACS sorting or 4×106 macrophage/well in 24-well plates for flow cytometry. Non-adherent cells were removed by washing after 3-5 hours, and macrophages were cultured in fresh RPMI-10 overnight. 400ng/mL purified E. coli O111:B4 LPS (List Biological Laboratories Inc.) was added to generate inflammatory macrophages 12h before the addition of target cells. …

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

4.1 | Reagent

… Purified LPS was purchased from List Biological Lab …

4.6 | Western blot analysis

Western blots were performed essentially in the same manner as previously described.[22] Briefly, cells (0.5 × 106 cells/ml) were seeded into six‐well plates, incubated for 48 h, and pretreated with different concentrations (50 and 100 nM) of EXM for 1 h. These cells were stimulated with LPS (10 ng/ml) or poly[I:C] (10 μg/ml) for 18 h, col- lected, and washed with ice‐cold phosphate‐buffered saline (PBS). …

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/

… Additionally, mice received 400 ng pertussis toxin (PTX) (List Biological Laboratories) intraperitoneally (i.p.) on days 0 and 2 after 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)

… goat anti-cholera toxin B (CTB, 1:1000, List Labs …

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

… Pertussis Toxin (PTX) List Biological Laboratories, Kalifornien (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)

… For induction of active EAE, C57BL/6 mice were immunized s.c. at the tail basis with 50 µg MOG35-55 peptide (GenScript) emulsified in Complete Freunds adjuvant (CFA, BD) along with 100 ng pertussis toxin (PTX, List-BiologicalLaboratories) administration i.p. on day 0 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)