Bacterial Toxin Research Citations

Flow cytometry and cell sorting

p63-EGFP knock-in hiPSCs were differentiated for 10 days or 6 weeks, dissociated with Accutase (Thermo Fisher Scientific) for 60–90 min at 37 °C, harvested through a 40 μm cell strainer (BD Biosciences, San Diego, CA), and collected into keratinocyte culture medium (KCM; a 3:1 mix of DMEM without glutamine and Nutrient Mixture F-12 Ham; Thermo Fisher Scientific) supplemented with 5% fetal bovine serum (Japan Bio Serum, Hiroshima, Japan), 0.4 μg/mL hydrocortisone succinate (Wako), 2 nM 3,3′,5-triiodo-l-thyronine sodium salt (MP Biomedicals, Santa Ana, CA), 1 nM cholera toxin (List Biological Laboratory, Campbell, CA), 2.25 μg/mL bovine transferrin HOLO form (Thermo Fisher Scientific), 2 mM L-glutamine, 0.5% insulin transferrin selenium (Thermo Fisher Scientific), and 1% penicillin–streptomycin. Cells were washed in phosphate-buffered saline. The 1383D2 cell line was used as an EGFP (enhanced green fluorescent protein) negative control. Cells were sorted on a SH800 Cell Sorter (Sony Biotechnology Inc., Tokyo, Japan) and the data were analyzed using SH800 software.

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

Preparations

… The preparation of the keratinocyte culture medium (KCM) is as follows; Dulbecco’s modified Eagle’s medium (DMEM; Sigma aldrich, MO, USA) and F-12 medium (Sigma aldrich) were mixed at a 3:1 ratio. The medium was supplemented with 3.3 mM l-glutamine solution (Sigma Aldrich), 84 μM gentamicin (Merck Sharp and Dohme, NJ, USA), 0.3 μM amphotericin B (Bristol-Myers Squibb, NY, USA), 0.25 μM hydrocortisone (Kowa, Nagoya, Japan), 140 mU/mL insulin (Lilly, IN, USA), 2.0 nM 3,3′,5-Triiodo-l-thyronine (MP Biomedicals, CA, USA), 1.0 nM cholera toxin (List Biological Laboratories, CA, USA), …

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

Cell culture and stimulation

CA-Mφ were generated by priming BMM with 150 U/ml IFN-γ (R&D Systems, Minneapolis, MN) for 6 h, and then stimulating cells overnight with 150 U/ml IFN-γ and 100 ng/ml LPS (from Salmonella typhimurium; no. 225; List Biological Laboratories, Campbell, CA). …

• Product #225 – LPS from Salmonella typhimurium

Cell lines

… MCF-10A cell lines were maintained in the full growth medium, which consisted of DMEM/F12 (1:1) (Cat#11330-032, Gibco) supplemented with 5% horse serum (Cat#16050-122,
Invitrogen), 10 mg/ml insulin (Cat#12878-44, Nacalai Tesque), 0.5 mg/ml hydrocortisone (Cat#1H-0888, Invitrogen), 100 ng/ml cholera toxin (Cat#101B, List Biological Laboratories), …

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

… Pertussis Toxin, List Biologicals, 180, …

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

… Pertussis fimbriae 2/3 and pertussis pertactin (69 kDa protein) were purchased from List Biological Labs (Campbell, CA) …

• Product #186 – Fimbriae 2/3 from B. pertussis
• Product #187 – Pertactin from B. pertussis (69 kDa Protein)

Materials and methods

Jak2V617F transgenic (TG) mice were generated as previously described [12]. CD11b-diphtheria toxin receptor (DTR) TG mice were purchased from The Jackson Laboratory (Bar Harbor, ME, USA) [13]. These mice were crossed to generate Jak2V617F/CD11b-DTR compound TG. To deplete monocytes in vivo, recipient mice transplanted with BMCs from CD11b-DTR TG mice or Jak2V617F/CD11b-DTR TG mice were administered 15 ng/g diphtheria toxin (DT) (List Biological Labs, Campbell, CA, USA) by intraperitoneal injection every 2 days for 8 weeks. …

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

… Diphtheria toxoid (List Biological Laboratories, CA), …

• Product #151 – Diphtheria Toxoid, from Corynebacterium diphtheriae

Active-induced EAE Models (aEAE) and adoptive-transferred EAE (tEAE) models

… At 0 and 24 hours after immunization, mice were intraperitoneally injected with pertussis toxin (250 ng per mice, List Biological, Campbell, CA, 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)

Key Resources Table
Diphtheria toxin (DT) List Biological Laboratories Cat# 150

In Vivo Cell Depletion

To ablate specific cells, CCR2-DTR, CCR2-DTR, Rag2-/-Il2rg-/-, ROSA26-iDTRMrp8-Cre, BDCA2-DTR, and non-transgenic littermate controls were injected i.p. with 10 ng/g body weight DT on Day -1, Day 0, and Day +2 pi (Espinosa et al., 2017, Swiecki et al., 2010), unless noted otherwise.

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