Pertussis toxin ptx

The dose of drugs and injection routes are indicated in figure legends. ANP, BNP, CNP, SST, and OCT were supplied from GenScript USA Inc (Piscataway, NJ). GRP_18-27, NMB and ANP-4–23 were from Bachem (King of Prussia, PA). Histamine, chloroquine, Npr1 siRNA, Npr2 siRNA, Npr3 siRNA, and scrambled control siRNA were purchased from Sigma (St. Louis, MO). BNP-saporin (BNP-sap) and blank-saporin were made by Advanced Targeting Systems. Pertussis toxin (PTX) and gallein were from R&D Systems (Minneapolis, MN) or Selleck (Houston, TX). AP 811 was from Tocris (Minneapolis, MN). U73122 was from Selleck (Houston, TX).

For establishing relapsing-remitting EAE model, female SJL/J mice were immunized subcutaneously with 0.2 mL of emulsion containing 150 μg of myelin proteolipid protein139-151 (PLP_139−151, HSLGKWL0GHPDKF; HPLC-purity >98%) (ChinaPeptides, Shanghai, China) dissolved in 0.1 mL sterile phosphate buffered saline (PBS) and 0.1 mL complete Freund's adjuvant (CFA) (Chondrex, Redmond, WA, USA) supplemented with 4 mg/mL Mycobacterium tuberculosis (Sands et al., 2014 (link); Rovituso et al., 2016 (link)). These injections were distributed over two spots upper abdomen with 0.1 mL emulsion per site. Each mouse received an additional 200 ng pertussis toxin (PTX) (R&D Systems, MN, USA) diluted in 200 μL PBS by intraperitoneal injection on day 0 and day 2 post-immunization (p.i.). Clinical scores were calculated blindly by two researchers daily according to a 0–5 scale as follows (Kono et al., 1988 (link)): 1 = flaccid tail; 2 = moderate hind or front leg weakness; 3 = severe hind or front leg weakness; 4 = complete paralysis of limb; 5 = death.