Citations

Bacterial strains, toxin, and antibodies:

…Purified V. cholerae type Inaba 569B (catalog no. 100B), heat-labile (catalog no. 165B), Shiga (catalog no. 161), and diphtheria (catalog no. 150) toxins were purchased from List Biological Laboratories (Campbell, CA). …

List Catalog #165B has been discontinued.

RTCA system:

…The toxin concentration for a sample was quantified using a calibration curve as described previously, with minor modifications (36). In brief, a series of 2-fold dilutions (from 3.5 pg/ml to 1.8 ng/ml) of purified CT were made in DPBS with 0.1% BSA. …

Cell morphological analysis:

The Y-1 cells were viewed at a time point at which a significant CI change was detected during the continuous monitoring by the RTCA system. The real-time status of the Y-1 cells in the 96-well microplate incubated with purified CT was monitored with an inverted phase-contrast microscope and photographed with an integrated digital camera…

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae
• Product #150 – Diphtheria Toxin, Unnicked, fromCorynebacterium diphtheriae
• Product #161 – Shiga Toxin 1 from Escherichia coli

Adult ablation of MCH neurons improves glucose tolerance, even in absence of MCH signaling:

MCH neurons increase their firing rate in situ when extracellular glucose increases (Burdakov et
al., 2005). Ten days after DT injection, MCH-neuron-ablated mice showed reduced area-under
the-curve (AUC) during a glucose tolerance test … Hypothesizing that this phenotype of MCH-neuron
ablation resulted from the loss of a non-MCH neurotransmitter in MCH neurons, I measured
glucose tolerance in homozygous PmchDTR/DTR mice injected with DT.

… Some vendors have proven more reliable than others at producing DT that is free of side effects in wild-type mice. We have had good luck with DT from List Biological Laboratories, but problems with that obtained from Sigma-Aldrich. DT comes lyophilized in vials of 1 mg. …

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

Diphtheria toxin-induced monocyte depletion mouse model:

… CDllb-DTR and C57Bl/ 6 WT mice were injected i.p. with 5001..11 of diphtheria toxin (List Biological
Laboratories, Inc.) at a concentration of 21..1g/ ml the day before pump insertion. …

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

Transganglionic tracer cholera toxin subunit B (CTB) was used to label the axons retrogradely (Wan et al., 1982, Trojanowski et al., 1982). Six weeks after the dorsal column transection and the first sciatic nerve injection, the left sciatic nerve was exposed at hip level (above the treatment site) and 5 µl of 1% CTB (List Biological Laboratories Inc, Campbell, USA) was slowly injected into the left sciatic nerve via a Hamilton syringe with 31 gauge needle. …

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

… of comparative anatomy, a corresponding site on the rat is situated between the second and third metacarpal bone of the forefoot planta pedis (figure 1). Under anaesthesia with ether (Beijing Chemical Plant, Beijing, China), a total of 6 µL 1% CTB (List Biological Labs, Campbell …

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

Blood Sampling and Immunization:

Two blood samples were collected from infants to assess cortisol levels and immune responses to an immunization. The first sample was collected when infants were 5.75 months of age, and the second at 8.75 months of age. For all blood samples, animals were captured in their home cages and 1.5 ml of blood was drawn from the femoral vein of each infant between 9:0010:00 am on a day when behavioral data were not being collected. On some occasions multiple animals in the same cage had samples collected, thus for all samples the disturbance time (i.e. the time from cage entry to blood sample completion) was recorded (mean = 10.8 minutes, range: 4.28 27.03 minutes) to later control statistically for the effects of cage disturbance, capture, and venipuncture on hormone levels. Immediately following the blood draw at 5.75 months of age infants received an immunization with 0.1 mg of cholera toxin B subunit (List Biological Laboratories, Campbell, CA) subcutaneously between the shoulder blades. Blood samples were drawn into 3 ml syringes and transferred to sterile tubes and allowed to clot at room temperature. Serum was then extracted and stored at 80°C until assay.

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

Muscle afferent tracing:

As a way to assess completeness of deafferentation, 7 days after DRR, we made bilateral injections of the transganglionic tracer cholera toxin B subunit (CTB; List Biological Laboratories, 1% in distilled water) into the surgically exposed extensor carpi radialis (ECR) or biceps brachii muscles to label muscle afferents. A volume of 10 µl was slowly injected into multiple intramuscular locations using a Hamilton syringe (24-gauge). We have optimized the CTB injections to maximally label the motoneurons of injected muscles, taking advantage of recent findings that many forelimb motor pools have an extended rostrocaudal distribution…

Spinal tissue preparation and CTB staining:

… All staining was conducted on free-floating section, and sections were mounted on gelatin-coated slides. Sections of C3 to T1 spinal cord were processed for CTB labeling.

Immunofluorescence staining of CTB has been described previously (Tan et al., 2012). Briefly, free-floating sections were incubated at room temperature (RT) in PBS containing goat anti-CTB primary antibody (1: 1000; List Biologicals) in blocking buffer…

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt
• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

Retrograde tracer experiments:

Injections of the retrograde tracer cholera toxin B subunit (CTB from List Biological Laboratories, Campbell, CA) were targeted to the medial preoptic area (n = 6) and the arcuate nucleus (n=6), unilaterally…Glass micropipettes of 15-20µm internal diameter were filled with 0.25% CTB dissolved in 0.1 M phosphate buffer at pH 7.4 (PB) and lowered to the following stereotaxic coordinates…Once the pipette was in place, the CTB was injected by iontophoresis using a constant current source…

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

Cell culture and treatments:

Human bronchial epithelial (HBE1) cells (generously provided by Dr. Reen Wu at the University of California, Davis [27]) were cultured at 37 C in 95% humidified air containing 5% CO₂ using Dulbecco’s Modified Eagle’s Medium (DMEM/F-12) supplemented with 50 U/ml penicillin, 50 g/ml streptomycin, 10 ng/ml cholera toxin (List Biological Laboratories, Inc.), …

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

Materials:

The following variants of LPS were used in these experiments: Salmonella minnesota R595 (Re) from List Biological Laboratories, Inc., cat # 304; …

• Product #304 – LPS from Salmonella minnesota R595 (Re)