Citations

Differentiation of SVF into adipocytes

When 100% confluency was reached, SVF cells were induced with the adipogenic cocktail containing 1 μM dexamethasone (Sigma-Aldrich, D4902), 10 μg ml−1 insulin (Sigma, I9278) and 0.5 mM isobutylmethylxanthine (Sigma, I7018) supplemented in DMEM/F12 for 2 days, followed by incubation in the adipocyte culture medium (DMEM supplied with 10% FBS) containing 10 μg ml−1 insulin for another 2 days. Then the cells were maintained in DMEM supplied with 10% FBS for 2 days. For treatment with inhibitors, cells were fasted for 12 h and pretreated with gallein (10 μM; MedChemExpress, HY-D0254), Y-27632 (10 μM; Solarbio, IY0050), SB203580 (10 μM; MedChemExpress, 152121-47-6), PTX (100 ng ml−1; List Biological Laboratories, 180) or H89 (20 μM; MedChemExpress, HY-15979A) for 4, 1, 1, 12 or 1 h, respectively, before treatment with 20 mM sodium L-lactate.

• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer

Reagents. LPS from E. coli serotype 0111:B4 was purchased from List Biological Laboratories (Campbell, CA). …

Chromatin Preparation and ChIP Assay.

Macrophages were stimulated in the presence or absence of LPS (10 ng/mL) in RPMI containing 10% FBS for up to 8 hours. …

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

Establishment of the EAE model

According to a previously established method30, WT C57BL/6 mice were subcutaneously immunized with 200 μg of murine myelin oligodendrocyte glycoprotein (MOG)35–55 peptide (Catalog Number: MOG-001, Chinese Peptide Co., Ltd., Hangzhou, China) in complete Freund’s adjuvant (Catalog Number: 7001, Chondrex Inc., Woodinville, WA, USA). Each mouse was intraperitoneally injected with 400 ng of pertussis toxin (Catalog Number: 181, List Biological Laboratories, Campbell, CA, USA) twice at 0 h and 48 h postimmunization. …

• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)

Retrograde viral tracing from the trachea and the lungs
… In the subsequent confirmatory experiments, the Cre-dependent viral tracer, AAVrg-CAG-FLEX-tdTomato-WPRE10GH, was injected into the trachea of heterozygous CGRP-Cre mice (n = 2). Again, animals were anaesthetized with isoflurane (4% for induction and 1.8–2.5% for maintenance), a midline incision was made in the neck and the muscles overlying the trachea were retracted to expose the trachea. Next using a Hamilton syringe, 1 μl of AAVrg-CAG-FLEX-tdTomato-WPRE10GH together with 2μl of saline was injected into the lumen of the trachea. After 4 weeks, the animals were euthanized and their brains, trachea and vagal ganglia were collected for immunohistochemistry. Alternatively, cholera toxin B (CTB, catalogue number #104, List Biological Labs, 0.3% w/v, 5 μl per injection), a retrograde tracer, was injected into the trachea of Phox2b-Cre;loxP-ChR2 mice (n = 6) following the experimental procedures described above. After one week, the animals were euthanized and their brains and vagal ganglia were collected for immunohistochemistry.

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