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Corticospinal projection neuron and callosal projection neuron FACS-purification
Corticospinal projection neurons and callosal projection neurons were purified from C57BL/6J mice as previously described.49,77 Briefly, corticospinal projection neurons were retrogradely labeled at P1 from the cerebral peduncle by injection of cholera toxin B subunit (CTB) from Vibrio cholerae with FITC conjugate (List Biological Laboratories, Campbell, CA) under ultrasound microscopic guidance. At P2, neuron bodies in the motor cortex were isolated by microdissection of deep cortical layers, and dissociated to a single cell suspension by papain digestion and mechanical trituration. Callosal projection neurons were retrogradely labeled on P1 by injection of CTB into the contralateral hemisphere under ultrasound microscopic guidance. ...
... FoxP3DTR mice to receive transferred Treg cells were treated with 50 mg/kg diphtheria toxin (DT) (List Labs, 150) starting from 5 days prior to tumor inoculation and for every 3 days until time of euthanization. …
Characterization of primary NSCs and C17.2 cells
P3 generation NSCs were seeded into petri dishes t a density of 1×106/ml and cultured overnight. Then, the cells were either treated with different concentrations of BHB for 1 day or pretreated with 500 ng/ml PTX (List Biological, 181), ...
EAE induction and scoring
WT NOD and IgH[MOG] mice were immunized subcutaneously with 200 µg MOG[35-55] (synthesized at the CHU de Québec), emulsified in incomplete Freund's adjuvant (BD Difco) that was supplemented with 500 µg M. tuberculosis extract (BD Difco). On day 0 and day 2 post-immunization, mice received 200 ng pertussis toxin (List Biological Laboratories) intraperitoneally. ...
Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
Biological materials.
E. coli 055 : B5 LPS (Lot # 20318 A4) was obtained from List Biologicals. All data presented in this study were generated with the same batch of E. coli 055 : B5 LPS. The synthetic lipid A derivatives were reconstituted as 2 mg/mL in 90 % THF/10 % water. Stock solutions in small aliquots were stored at −80 °C.
EAE
Active immunization was induced by s.c. injection of MOG[35-55]11 (CHU de Québec) in incomplete Freund's adjuvant (Difco) supplemented with 5 mg mL-1 Mycobacterium tuberculosis extract (Fisher). A dose of 25 μg MOG[35-55] per mouse was used to compare WT to Serpine1-KO mice, while 100 μg per mouse was used in EAE experiments involving Tim-3-Tg mice. In Figure 1D, passive EAE was induced by first immunizing Serpine1-WT 2D2 and Serpine1-KO 2D2 mice with MOG[35-55]. Nine days later, splenocytes were collected and stimulated ex vivo for 48 hours with MOG[35-55] (20 μg mL-1), rmIL-12 and rmIL-23. Next, 20x106 splenocytes were injected i.p. into unimmunized B6 recipients. Adoptive transfer of WT 2D2, Serpine1-KO:2D2 or RV-transduced 2D2 Th1 cells entailed i.v. injection (2x106) into Rag1-/- recipients at d5 of culture. In all EAE experiments, mice received 200 ng pertussis toxin (List Biological Labs) i.p. at d0 and d2. Mice were assessed for clinical symptoms daily as previously described (9).
Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
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