Bacterial Toxin Research Citations

Mice and Treg depletion:

… To deplete Treg cells, DEREG mice were injected with DT (List Biological Laboratories). Mice were injected i.p. with 1 mg DT every other day over a period of 9 d starting with the rst injection 1 d prior MHVA59 infection. For adoptive transfer experiments, recipients were treated with DT on days 5, 7, and 9 postinfection (p.i.). Control mice (B6) received the same DT treatment as DEREG mice. 2D2 3 DEREG donor mice were injected with 1 mg DT 1 d prior to cell isolation.

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

Animals and Drug Treatments:

Diphtheria toxin (DT; List Biological Laboratories) was administered intraperitoneally at 15 ng/g, on 2 consecutive days.

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

Animal maintenance and neuron ablation

… All except Tph2^lox/lox mice were on the C57Bl/6 background (> 9 generations backcrossed); Tph2^lox/lox mice were on a mixed 129/Sv x C57Bl/6 background. Agrp^DTR/DTR male mice were bred with Slc17a6^lox/lox female mice to generate Agrp^DTR/+; Slc17a6^lox/+ mice, which were further bred to each other to create Agrp^DTR/+; Slc17a6^lox/lox mice and Agrp^DTR/+; Slc17a6^lox/+ control mice. Similar breeding strategy was adopted when generating Agrp^DTR/+; Tph2^lox/lox mice and Agrp^DTR/+; Grin1^lox/lox mice. Mice were group housed with standard chow diet (LabDiet 5053) and water provided ad libitum until the beginning of the experiments. All experiments were performed with ~8-wk-old male mice. To ablate AgRP neurons, mice carrying the Agrp^DTR allele, were injected twice with DT (50 g/kg; intramuscular; 2 d apart; List Biological Laboratories, Campbell, CA)⁴. The extent of ablation (>95%) was determined by immunohistochemistry⁴⁷.

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

Mice:

C57BL/6 (CD45.2⁺) mice and congenic C57BL/6 (CD45.1⁺) mice were bred in and provided by the animal facility of the Max Planck Institute of Immunobiology and Epigenetics, Freiburg, Germany. … Cell ablation was achieved by a single i.p. injection of DT (30 ng/g body mass) (List Biological Laboratories) unless stated otherwise. Note that, after birth, mice heterozygous or homozygous for the Ccx-Ckr1-eGFP knock-in allele have no overt thymus phenotype (7).

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

Mice and Reagents:

Wild-type C57BL/6 mice were purchased from Jackson Labs. … Control mice received PBS. In vitro stimulation of single-cell suspensions from the lamina propria was performed with a concentration of 200 ng/ml of flagellin. Diphtheria toxin (List Biological Laboratories) was administered every other day at a dose of 10 ng/g starting 6 days prior to injection of flagellin. Mice received a total of three doses.

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

Mice and Reagents:

Wild-type C57BL/6 mice were purchased from Jackson Labs. … Control mice received PBS. In vitro stimulation of single-cell suspensions from the lamina propria was performed using a concentration of 200 ng/mL of flagellin. Diphtheria toxin (List Biological Laboratories) was administered every other day at a dose of 10 ng/g starting 6 days prior to injection of flagellin. Mice received a total of 3 doses.

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

Animals and DT Treatment:

…Before behavioral experiments, mice were maintained on standard laboratory chow (5053; Lab Diet) that was available ad libitum. When the Lep^ob/ob;Agrp^DTR/+ and Lep^ob/ob;Pmch^DTR/+ mice were 6 wk old (weighing 2025 g), 8 wk old (weighing 3540 g), or 10 wk old (weighing 4045 g), they were individually housed and switched to another standard chow diet (D12450B; Research Diets) for 7 d before being injected with DT. To ablate AgRP or MCH neurons, DT was injected twice per mouse (i.m., 2 d apart; List Biologicals). Body weight and food intake were recorded daily for several weeks. The DT dose was determined based on body weight at the time of injection, with 50 g/kg for mice weighing <40 g and 40 g/kg for mice weighing >40 g. …

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

Regulatory T cell transfer and Foxp3+ T cell ablation:

Spleens and lymph nodes from either WT or Foxp3DTR B6 3 B6.g7 mice were harvested, and polyclonal CD4+ T cells were puried by negative selection using the CD4+ T cell isolation kit and an LS column (Miltenyi Biotec). To deplete Foxp3-expressing cells from reconstituting polyclonal Foxp3DTR CD4+ T cell populations, DT (List Biological, Campbell, CA) was injected i.p. at a concentration of 50 mg/kg body weight on days 8, 10, and 12 after CD4+ T cell reconstitution (19). …

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

DC and NK Cell Depletion:

Depletion of cDCs was induced in CD11c-DTR-Gfp mice as described.35 Briefly, 100 ng (4 ng/g) of diphtheria toxin (DT) (List Biological Laboratories Inc.) was administered intraperitoneally 1 day after the last dose of CCl₄. Three days later, the mice were sacrificed and livers were harvested (Supporting Fig. 2B). NK cell depletion using anti-asialo-GM1 antibody (Wako Chemicals) and NKp46-DTR-eGFP mice were performed as described32, 36 (Supporting Fig. 2F,G).

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