Citations

Immunohistochemistry:

… Some of the brains (n=3) from WT mice were injected with CTb and were immunostained using Goat anti CTb (1:30K, Cat# 703, RRID: AB_10013220, List Biological Laboratories Inc., CA). Sections for double staining for GFP, mcherry, CTb or FoxP2 were incubated in fluorescent-labeled secondary antibodies (Alexa- 488 at 1:200 or Alexa- Cy3 at 1:200; …

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

2.2. Wt-RABV injections in the inferior live of the rat (n = 10)

The wt-RABV stock was kept at −80◦C until use. The injectate consisted of a mixture of 4 parts wt-RABV and 1 part cholera
toxin β-subunit [CTb, low salt; List Biological Laboratories, 1% w/v in 0.2 M phosphate buffer (PB) at pH 7.4]. CTb was added to accurately evaluate the injection site and to be able to discriminate between non-transneuronal (CTb) and non-transneuronal and transneuronal infection by the wt-RABV (Ruigrok et al., 2008; Suzuki et al., 2012). Injection of approximately 150 nl was made with a 1 µl Hamilton needle attached to a microsyringe pump (Ruigrok et al., 2016). The IO was reached at a −45◦ angle with the vertical axis, with the needle entering at 0.6–0.8 mm lateral to obex at a depth of 2.7–2.8 mm. After injection the neck muscles were sutured in layers, the skin was clamped and the animal was allowed to recover. …

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

… Prediluted serum samples (1:50) were diluted by 2-fold in another 96-well plate and incubated with a constant amount of anthrax lethal toxin (100 ng/mL PA and 80 ng/mL LF; List Biological Laboratories, Campbell, CA, USA) for 30 min. The generated toxin-antiserum was then applied to J774A.1 cells and incubated for 4 h. …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

Stereotaxic surgery.

… For retrograde tracing of TRN-projecting and PVT-projecting PL cells CTB-488 and CTB-555 (1.0% in PBS, Thermo Fisher Scientific) were injected into the TRN (0.5 µL) and PVT (1.0 µL), respectively, and allowed 4 d for retrograde transport. For retrograde tracing of PVT-projecting TRN cells unconjugated CTB (List Labs Product No. 104) was injected into the PVT (1.0 µL). For retrograde labeling of PVT-projecting TRN cells for RNAscope, retrobeads (LumaFluor, Inc.) were injected into the PVT and allowed 7 d for retrograde transport. …

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

… immunohistochemistry. Briefly, sections … were processed and incubated overnight in a goat anti-CTb antibody (List Biological Lab, #7032A6) at 1:10,000, followed by rinses in PBS and incubation …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

Induction of EAU

To effectively induce EAU production, we injected mice subcutaneously with a 1:1 mixture of 200 µg IRBP1−20 (GL Biochem, Shanghai, P. R. China) and complete Freund’s adjuvant (Difco, Detroit, MI, USA) including 2.5 mg/mL of Mycobacterium tuberculosis strain H37Ra (BD Difco, San Jose, CA, USA). After immunization, 0.25 μg of PTX (List Biological Laboratories, Campbell, California, USA) was injected intraperitoneally 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)

Active EAE and adoptive transfer EAE

In the active EAE model, FACS-sorted MP CD4+ T cells (γδTCR–NK1.1–Vβ11–TCRβ+CD4+CD1d tetramer–Foxp3–CD62LlowCD44high, 5 × 105) from Foxp3-GFP mice were adoptively transferred into 5-week-old female C57BL/6 mice. After the transfer, the mice were immunized with 200 μg of MOG35-55 peptide in complete Freund’s adjuvant (Chondrex, Inc., Woodinville, WA, USA). At 0 and 48 h after immunization, the mice were intraperitoneally treated with 500 ng of pertussis toxin (List Biological Laboratories, Inc., Campbell, CA, USA). The animals were scored daily for clinical disease.

In another EAE model established by adoptive transfer, naive (CD4+Vβ11+CD25–CD62LhighCD44low) CD45.1− T cells (1–5 × 104) from 2D2 TCR-transgenic mice were transferred into Rag–/– mice with or without WT CCR6high, WT CCR6low, Bhlhe40–/– CCR6high, GM-CSF–/– CCR6high, GM-CSF–/– CCR6low, Il1r1–/– CCR6high or Il1r1–/– CCR6low MP CD4+ T cells (CD45.1+γδTCR–NK1.1–Vβ11–TCRβ+CD4+CD1d tetramer–CD25–CD62Llow CD44high, 1.0 × 105). Before the transfer, CCR6high or CCR6low MP CD4+ T cells were primed in vitro with IL-7 (10 ng/mL) and IL-1β (20 ng/mL) for 5–7 days. After the transfer, the mice were immunized with 100 μg of MOG35−55 peptide in complete Freund’s adjuvant. At 0 and 48 h after immunization, the mice were intraperitoneally injected with 200 ng of pertussis toxin (List Biological Laboratories, Inc., Campbell, CA, USA). …

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)

… CD11c-DTR MRL/lpr mice and transgene-negative littermate controls were i.p. injected with a single dose of DT (List Biological Labs) at a concentration of 8 ng/kg body weight in PBS at 1, 4, or 7 d prior to experimental analysis …

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

… Additionally, mice received intraperitoneal injection of pertussis toxin (List Biological Laboratories, Campbell, California, USA) on the same day and the second day after the immunization. …

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)

Lipopolysaccharide murine acute lung injury model

… Eight- to ten-week-old C57BL/6 J female mice (Jackson Laboratory) were injured intratracheally (IT) with lipopolysaccharide (LPS, O55:B5 #203A, List Labs). Each mouse received 50 μg LPS dissolved in 50 μL sterile phosphate buffered saline (PBS) at time zero. Control mice were administered 50 μL IT PBS. …

• Product #203A – LPS from Escherichia coli O55:B5