Bacterial Toxin Research Citations

STAR★Methods
Key resources table

… Diphteria toxin, List biological Labs …

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

… DT (List Labs, cat. no. 150) was dissolved in 1× PBS and administered by intraperitoneal injection …

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

In vivo adeno-associated virus delivery

… To reduce DP macrophages, diphtheria toxin (DT) (List Biological Laboratories) was diluted in PBS and administered to mice IP acutely (25 ng/mg body weight) or chronically (25 ng/mg every other day for 4 weeks).

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

A single dose of 200 ng DT (List Laboratories, #150) was intraperitoneally introduced one week after the last Tamoxifen injection. The cancer cells were inoculated into

amoxifen and diphtheria toxin treatment

Tamoxifen (MedChemExpress, #HY-13757A) was dissolved in corn oil (20 mg ml−1, Wako, #032-17016) and heated at 37 °C for 3 hours. Mice were administered with tamoxifen through oral gavage (100 mg kg−1 body weight). To activate Cx3cr1CreERT2-mediated recombinase, each mouse was fed with Tamoxifen every three days, starting from Day 12 post-intraportal injection, as indicated in Fig. 2e. For proliferation tracing experiment, DreERT2-mediated recombinase was introduced by a single dose of tamoxifen on Day 12 post-intraportal injection, as indicated in Fig. 3f. For KC specific depletion, DreERT2-mediated recombinase was introduced by 8 doses of tamoxifen gavage, as indicated in Fig. 5c. A single dose of 200 ng DT (List Laboratories, #150) was intraperitoneally introduced one week after the last Tamoxifen injection. The cancer cells were inoculated into liver through portal vein one day after DT injection.

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

… Diphtheria toxin (DT) List Biological Laboratories Cat#150 …

In vivo Cell Depletion

To ablate specific cells, BDCA2-DTRTg/+, BDCA2-DTRTg/+ Stat1−/−, 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 (Swiecki et al., 2010), unless noted otherwise.

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

CCR2+ inflammatory monocyte and CD11c+ cell depletion

The CCR2 depleter (CCR2-DTR) and CCR2 reporter (CCR2-GFP) strains were generated on the C57BL/6 (WTB6) background as previously described (48). Control animals for CCR2-DTR experiments were sex and age-matched, nontransgenic littermates. CD11c-DTR mice in C57BL/6 background were from Dr. Kumamoto’s Laboratory at Rutgers University. CD11c-DTR mice were generated with a targeting construct containing a floxed DTR-IRES-GFP cassette containing a splice acceptor that was knocked into the first intron of the CD11c gene, known as integrin (Itgax) (62). For depletion of CCR2+ or CD11c+ cells, CCR2-DTR mice and control CCR2-DTR negative littermates; or CD11c-DTR mice and their littermate controls, received 250 ng of diphtheria toxin i.p. 1 day prior to infection and every other day thereafter in order to maintain depletion. Diphtheria Toxin was purchased from List Biological Laboratories (Campbell, CA), and reconstituted at 1 mg/mL in PBS.

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

… Diptheria toxin List Biological Laboratories Cat#150 …

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

Mice

Rag2−/− (Cat# 033562), Rag2−/−γC−/− (Cat# 014593), Ccl2−/− (Cat# 004434), Foxp3YFP-Cre (Cat# 016959), Scurfy mice (Foxp3−/y, Cat# 006775), Foxp3DTR knock-in mice (Cat# 016958) were purchased from The Jackson Laboratory (Bar Harbour, USA). Ccr2cKO mice were generated by crossing Ccr2loxP (Gempharmatech, Nanjing, China) mice with Foxp3YFP-Cre mice. For Tregs ablation, 6-week-old Foxp3DTR mice were intraperitoneally injected twice a week with 500 ng dose−1 of diphtheria toxin (DT, List Labs, California, USA) for 3 weeks. All mice were housed and maintained under specific pathogen-free (SPF) conditions. …

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

… For cell ablation, 1mg/ml stock solution of DTX (#150, List Biological Laboratories) was prepared with water, aliquoted and stored at 20°C until use. When needed, the DTX solution was diluted to 5ng/µl with sterile 0.9% saline and 50µg/kg was administered intraperitoneally (i.p.) daily for 5 consecutive days. …

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

… Diphtheria Toxin from Corynebacterium Diphtheria (unnicked) List Biological Laboratories Inc Cat# 150 …

Tissue dissection, islet isolation and cell culture

… Islets and myeloid fractions obtained from each pancreas were combined by overnight culture in AggreWells (Stem Cells Technology) in RPMI-10% FCS-5mM glucose; recovered islets/myeloid cell aggregates were then cultured up to 10 days in the same medium in the presence or absence of 50 ng/ml of DT. Culture medium and DT were replaced every 48 hours.

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