Citations

EAE induction

Mice were immunized subcutaneously with 100 µl of emulsified complete Freund’s adjuvant (CFA, Sigma) supplemented with 10 µg/µl Mycobacterium tuberculosis and 100 µg MOG35–55 peptide (Charité Berlin) and received intraperitoneal injections of 200 ng pertussis toxin (List Labs) at the time of immunization and 48 h 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)

… All mice received two subcutaneous injections of 100 µl each of the mix of MOG and complete Freunds adjuvant. All mice then received an intraperitoneal injection of Pertussis toxin (List Biological Laboratories, 180) …

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

STAR+METHODS
KEY RESOURCES TABLE

… LIST HPT LPS from E. coli O113 List Biological Laboratories …

• Product #433 – LIST™ HPT™ from Escherichia coli O113

… Cat# 555029 Diphtherin List Labs Cat# 150 …

Adoptive transfer of immune cells

For the adoptive transfer of BMDCs, Itgax(CD11c)-DTR/EGFP-recipient mice (age 6–7 weeks, male) were intraperitoneally injected with 100ng diphtherin (DT) and administered with WT BMDC or Slc38a5−/− BMDC cells (1 × 106 cells suspended in 100 μL sterile PBS) via the ophthalmic vein. DT injection and cell transfer were performed every 3 days starting from one day prior to IMQ treatment, and 3 times of DT injection plus cell transfer were performed (Figure S4B). The mice in the control groups received injection of sterile PBS (100 μL). …

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

… Mice aged 8-10 weeks were immunized subcutaneously with 100 µg of MOG35-55 peptide (MEVGWYRSPFSRVVHLYRNGK) emulsified in CFA supplemented with Mycobacterium tuberculosis H37Ra extract (Difco), followed by pertussis toxin (List Biological Laboratories) injections (150 ng per mouse) on days 0 and 2, as described previously …

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)

… The retrograde tracers Fluorogold (fg, 5-hydroxystabilamide; Cat No. 80014, Biotium, Hayward, CA, USA) and Cholera toxin b subunit (ctb, Cat No. 104, List Biological Inc., Cambel, CA, USA) using a 1µl Hamilton syringe …

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

… Pertussis toxin (List Biological Laboratories, Inc., Campbell, CA) was injected i.p. on the day of immunization and 48 h later. Weight and clinical scores were monitored every two 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)

Cell culture method

… For M1 polarized cells, on Day 3, PMA media is removed, and cells are washed with PBS. Thereafter, polarizing media consisting of complete media with LPS (List Biological Laboratories, 421, 100 ng/mL) and IFN-y (Sigma-Aldrich, SRP3093, 50 ng/mL) were added to the dish. The cells were incubated for another two days. On day 5, the media was removed, and cells were washed with PBS before fixing. …

• Product #421 – ULTRA PURE LPS from Escherichia coli O111:B4

MOG-induced EAE in / 6 mice

The mice were immunized by subcutaneous injections of 200 μg myelin oligodendrocyte glycoprotein (MOG) (35–55) (Genemed Synthesis, San Francisco, CA) in complete Freund’s adjuvant, containing 1 mg M. tuberculosis, strain H37RA (CFA; Difco, Detroit, MI), divided among two sites draining into the axillary lymph nodes. Sham mice, that served as negative controls, were injected with complete Freund’s adjuvant without MOG (35-55). After immunization (day 0) and on day 2, the mice received intraperitoneal injections of 200 ng pertussis toxin (List Biological Laboratories, Campbell, CA) dissolved in phosphate-buffered saline (PBS).

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)

Immunohistochemistry, image capture, and Imaris reconstruction

Mounted spinal cord sections were first incubated in PBS containing 0.1% Triton X (PBS-T) for 3 consecutive washes of 5 minutes each. Subsequently, spinal cord sections were incubated in 0.1% PBS-T solution containing primary antibodies and 10% heat-inactivated horse or donkey serum (Invitrogen) overnight at 4°C. Primary antibodies used were rabbit anti-DsRed (1:2000) [Clontech, 632496], guinea pig anti-vglut1 (1:1000) [Millipore, AB5905], goat anti-CTB (1:10 000) [List Biological Laboratories, 703], and chicken anti-parvalbumin (1:500) [sysy, 195006]. Following primary antibody incubation, spinal cord sections were washed with PBS for 15 mins (3x5min fresh solution). …

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