Citations

Reagents

Pertussis toxin was purchased from List Biological Labs (Campbell, CA).

EAE induction

For chronic EAE induction, female C57BL/6 mice with an age of 8–9 weeks old were injected subcutaneously with an emulsion of 200 μg MOG35–55 peptide in Complete Freund’s adjuvant containing Mycobacterium tuberculosis H37 RA (final concentration 2 mg/ml) on day 0 followed by intraperitoneal administration with 200 ng pertussis toxin on day 0 and day 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)

… The mice were then subsequently intraperitoneally injected with 200 ng of pertussis toxin (List Biological Laboratories) on day 0 and day 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)

Key resources table

Peptide, recombinant protein Staphylococcal enterotoxin B List Biological laboratories # 122 1 µg/ml, PBMC stimulation

… Full-length Clostridium difficile toxin A (Cat#152C) was purchased from List Biological (Campbell, CA) …

• Product #152C – Toxin A from Clostridium difficile

Establishment Of The Eae Model

… Pertussis toxin (PTX; List Biological Labs, Campbell, CA) (300 ng) was intraperitoneally administered on the rst and third days 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)

… Then, this emulsion was injected intravenously followed by intraperitoneal injection of 300 ng pertussis toxins (List Biological Lab, USA) at the day 0 and day 2. After EAE scoring reached one (approximately 10 days 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)

Experiment design to test for a circadian phase-dependent response to TcdB

1-2 HIOs and ≈15-20 mouse/human enteroids were plated per well to control for density.
100nM dexamethasone (DEX) was added to treatment groups either 36-hours (D36) or 24-hours
(D24) prior to TcdB addition. Samples were then exposed to a vehicle buffer (50mM Tris Base
Ultrapure (US biological), 100mM NaCl (Fisher Scientific), pH 7.5) or Clostridium difficile toxin B
(List Laboratories) by addition to cell culture media. …

• Product #155 – Toxin B from Clostridium difficile

Primary Cells Extracted from Human Corneal Biopsies

Td_hLESCs were collected from scleral‐corneal tissue (obtained with written consent from next of kin according to the directives set by the Italian Centro Regionale Trapianti and Centro Nazionale Trapianti, and only when not suitable for transplantation) and preserved in media before processing. Biopsies, composed of cornea, limbus, and conjunctiva, were dissected into quarters, and then the cornea and conjunctiva were removed. The limbus was fragmented and soaked in PBS. Trypsin/EDTA (0.05%/0.01% w/v; Thermo Fisher Scientific, Italy) was added (10 mL) to digest the tissue for 30 min at 37 °C. The solution was centrifuged for 5 min at 1000 rpm and the pellet resuspended in fresh media. This cycle was repeated thrice to extract the majority of the cells. Cells were plated into a 24‐well plate (30 000 cells per well) in coculture with irradiated 3T3‐J2 cells. Culture medium consisted of Dulbecco’s modified Eagle’s medium (DMEM) and Ham’s F12 (DMEM/F12, 2:1, Thermo Fisher Scientific) supplemented with 10% v/v FBS (Thermo Fisher Scientific), 50 µg mL–1 penicillin‐streptomycin (P/S), 4 × 10−3 m glutamine (EuroClone, Italy), 5 µg mL–1 insulin (HUMULIN R, Lilly, Canada), 0.4 µg mL–1 hydrocortisone (Flebocortid Richter, Sanofi, Italy), 0.18 × 10−3 m adenine (adenine grade I, Pharma Waldhof GmbH, Germany), 8.1 µg mL–1 cholera toxin (Cholera Toxin QD, List Biological Laboratories, USA), 2 × 10−3 m triiodothyronine (Liotir, IBSA, Italy), and 10 ng mL–1 epidermal growth factor (GMP Cellgro, CellGenix GmbH, Germany).

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

… For macrophage activation, the adherent cells were stimulated with 1g/mL of lipopolysaccharide from E.coli O55:BS (List Biological Lab Inc.) in media for 4 h at 37 C …

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

… Retrograde tracing was performed using a 1% solution of cholera toxin B subunit (CTB; List Biological Labs), injected using standard stereotaxic coordinates (Paxinos and Franklin, 2001). …

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