Bacterial Toxin Research Citations

… DT powder is purchased from Sigma-Aldrich #D0564 or List Biological Labs #150 and can be stored at 2-8 °C …

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

… For DC depletion, mice were injected with 500 ng of DT (List Biological Laboratories) intraperitoneally at indicated time points. …

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

Administration of Drugs

DT (15046A1, List Biologicals Laboratories) was used as previously described.26 Conditional ablation of Tregs in DEREG mice was achieved by intraperitoneal injection of DT (50 µg/kg body weight) 2 days before the MCAO model. Subsequently, an intraperitoneal dose of DT (10 µg/kg body weight) was administered every other day until day 3 after MCAO. …

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

Animal studies and treatments

… Mice were treated with 0.5µg DT or CRM197 (List Biological) i.p. on day 0 only or on days 0, 4, 7, and 10. For in vivo competition assays, mice were treated on day 0 with 0.125-0.5µg DT and/or CRM197. …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae
• Product #149A – Diphtheria Toxin CRM197, Mutant

… DT (List Biological Laboratories, 150) was dissolved in PBS, and 1µg (first dose) or 250ng (subsequent doses) was injected intraperitoneally for each mouse …

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

Allergic sensitization and mouse model of asthma

For allergic sensitization, mice were lightly anesthetized with isoflurane and given two oropharyngeal (o.p.) instillations, 1 week apart, of 100 μg LPS-free OVA (Worthington Biomedical) with 10 μL HDE or 100 ng LPS (Millipore Sigma) in a total volume of 50 μL in PBS (HDE/OVA or LPS/OVA)30. The HDE was prepared as previously described32,59. Briefly, vacuumed dust samples from homes in North Carolina were passed through a coarse sieve, then extracted at 100 mg/mL with PBS at 4 °C with overnight mild agitation. The samples were centrifuged to remove insoluble debris, and supernatants were sterilized by passage through a 0.22 μm filter (Millipore Sigma). Endotoxin concentration was 50 ng LPS/10 μL HDE, as measured by a Limulus Amebocyte Lysate assay (Lonza, catalog #50-648U). In some experiments, 100 μg Alexa Fluor (AF) 647-conjugated OVA or 10 μg DQ-OVA (ThermoFisher Scientific) was used to analyze antigen uptake60. In some experiments, DTX (20 ng/g body weight) (List Biological Laboratories) was injected into peritoneal cavity of zDC-DTA mice prior to airway sensitization …

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

2.1. Mouse model and treatment

Vglut3DTR/+ mice were generated in our lab. We used the CRISPR/Cas9 approach to connect DTR to the C-terminus of Vglut3 via the P2A peptide. PCR and Southern blot methods were used to screen out the mice with the correct recombination. Lgr5CreERT2/+ (Stock #008875) and Rosa26-tdTomato (Stock #007914) mice were purchased from the Jackson Laboratory. The primers used for genotyping Lgr5creERT2/+, Rosa26-tdTomato and Vglut3DTR/+ mice are listed in Supplementary table 1. Tamoxifen (Sigma-Aldrich) was injected intraperitoneally at postnatal day (P)2 at 0.20 mg/g bodyweight. EdU (Click-iT EdU Imaging Kit; Invitrogen) was injected intraperitoneally from P3 to P5 once a day at a dose of 50 μg/g. DT (List Biological Laboratories) was injected intraperitoneally at 12.5 ng/g, 25 ng/g, or 37.5 ng/g at P2. …

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

Diphtheria toxin (DT) administration.

Experimental mice (DTR mice, 6-9 weeks old) received 2 123 intramuscular injections of DT from Sigma-Aldrich (St. Louis, MO) or List Biological Laboratories, Inc. (Campbell, CA) at 25 ng/g, spaced 2d apart …

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

Diphtheria-toxin-induced depletion of TRMs.

Depletion of TRMs was induced by intranasal instillation of 15 ng per mouse of diptheria toxin (DT, List Biological Laboratories).

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

Ablation of AgRP neurons

To ablate AgRP neurons, mice carrying the AgrpDTR/+ allele, were bilaterally microinjected with DT (0.4 ng/side/mouse, List Biological Laboratories, Campbell, CA) using a needle (Hamilton Small Hub RN 33 G, Reno, NV) connected with a 10 μl syringe (Hamilton 700 Microliter, Reno, NV), at a rate of 0.1 μl/min. A total 0.4 μl volume/side was delivered into the dlDRN2. To describe the expression profile of AgRPARC-dlDRN neurons, the vehicle (0.9% NaCl) or DT were injected into the dlDRN with AgrpDTR/+::NpyGFP mice. …

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