Citations

… All dosing solutions were prepared in DMEM from stocks immediately prior to dosing for final dosing concentration of 40M or 10M and 0.0004% DMSO. Stocks were prepared of ultrapure lipopolysaccharide (LPS; 1 endotoxin unit (EU)/ng, E. coli 055:B5 Lot#4231A1 List Biological Laboratories, Campbell, CA) with sonication in nuclease free sterile water …

• Product #423 – ULTRA PURE LPS from Escherichia coli O55:B5

… On day 0, the mice were also administered 500 ng of pertussis toxin (List Biologicals, Campbell, CA, USA) intraperitoneally. …

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)

… a group of Wistar rats received unilateral microinjections of the retrograde tracer Cholera Toxin B subunit (CTB; 1% w/v in artificial cerebrospinal fluid (ACSF), 100 nL, List Biological Laboratories, Campbell, CA, USA) into the BLA (all coordinates are expressed in mm from bregma, AP: 1.9; ML: 5.00; DV: 8.5) in experiment 1 …

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

Bioassay Reagents

… Active CT produced in V. cholerae was purchased from List Biological Laboratories, azide-free in powder form (product no. 100B). …

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

EAE immunization and scoring

As described previously27,28,29,30, mice were immunized s.c. on d0 in the right and left flank with 100ug of PLP antigen emulsified 1:1 volume in complete Freund’s adjuvant supplemented with 4 mg/ml Mycobacterium tuberculosis (CFA; Becton Dickinson, Franklin Lakes, NJ), followed by 250 ng pertussis toxin (PTx; List Biologicals, Campbell, CA) i.p. on days 0 and 2. …

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)

EAE induction, treatment and assessment. Mice were immunized at multiple sites by subcutaneous injection of 200 μg of MOG35-55 or 200 μg of OVA protein (as a control) (Sigma-Aldrich) emulsified in 200 μl of complete Freund’s Adjuvant supplemented with 5 mg/mL heat-inactivated Mycobacterium tuberculosis H37Ra (Thermo Fisher). A total of 200 ng of pertussis toxin (List Biological Laboratories, Campbell, CA, USA) was injected intravenously into the mice after the MOG3555 injections and again 2 days later. …

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)

Unloading by BOTOX-induced paralysis

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12-week-old wild type and Sp7-Cre driven Piezo1/2 DKO male mice were anesthetized by isoflurane and then injected with 0.5U botulinum toxin (BTX, List Biological Laboratories) into both the right quadriceps and the right calf muscles. This treatment is a tibial unloading model by muscle paralysis and disuse. 24 hr after BTX injection, mice were unable to use their right hind limb. Left tibiae served as normal loading controls. Since a rapid and profound bone loss could be observed 1 week after BTX injection (Grimston et al., 2007), the mice were euthanized 10 days after injection and the tibiae were analyzed by μCT. A 0.8 mm thick section spanning the proximal tibia metaphysis was analyzed as shown in previous studies (Judex et al., 2004).

Macrophage exposures RAW264.7 cells were seeded into 24-well plates (200,000 cells/well). The next day, the medium was replaced with medium containing the indicated doses of either E. coli O111:B4 LPS (List Biological Labs), PAM3CSK4 (Invivogen), or poly(I:C) (Invivogen). Following the exposure, cells were lysed in RLT for RNA preparation. Additionally, in some studies, supernatant was collected for analysis of cytokine protein levels by ELISA. In the studies in which the cells were given a pulse of LPS, …

• Product #201 – LPS from Escherichia coli O111:B4

… PBMCs were either left unstimulated (DMSO) or stimulated for 6h at 37 C with either SIVmac239 overlapping Gag/Env peptides (NIH AIDS Reagent Program) at a concentration of 1gml1 or staphylococcal enterotoxin A & B (SEB/AList Biologicals) at 250ngml1 . …

Induction of EAE

EAE was induced using a modification of our previously reported method [15]. In brief, 8-week-old wild type or Sema4A-deficient C57BL/6 female mice were subcutaneously injected with 100 μg MOG35–55 emulsified in complete Freund’s adjuvant (CFA) supplemented with intraperitoneal injections of 200 ng pertussis toxin (List Laboratories, Campbell, CA, USA) on days 0 and 2. …

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)