Pf 05089771

From day 8 to day 11 after surgery, Nav1.7 channel blockers including PF-05089771 (Tocris, #5931, 2 mg/kg, and 4 mg/kg) and GNE-0439 (ProbeChem, # PC-62325, 10 μg/kg, 20 ug/kg, and 30 ug/kg) were administrated individually via IP injection into adult C57BL/6 SCI mice and mechanical pain threshold was tested at 30- and 60-min post drug administration. The voltage-gated calcium channel blocker Gabapentin (Sigma, #G154, 50 mg/kg) was administrated via IP injection into SCI mice and mechanical threshold was tested at 30- and 60-min post drug administration. Each drug test had a vehicle (saline or 20% DMSO) group and pre-treatment threshold was firstly measured each day.

ICA 121431 (#5066), 4,9-anhydrotetrodotoxin (AH-TTX, #6159), and PF-05089771 (#5931) were purchased from Tocris Bioscience. Tetrodotoxin (TTX, ab120054) was purchased from Abcam. All other chemicals were from Sigma-Aldrich and Fisher Chemical.

Normal goat serum and lysophosphatidic acid (LPA) were obtained from Abcam (Toronto, ON, Canada), PF05089771 from Tocris Bioscience (Toronto, ON, Canada), AM281 from Cayman Chemical Company (Ann Arbor, MI, USA), PEP-1 from Abbiotec Inc. (San Diego, CA, USA), Fluoro-gold from Fluorochrome LLC (Denver, CO, USA), mouse IgG from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA), and guinea pig anti-Na_v1.7 (for IHC) from Alomone Labs (Jerusalem, Israel).
The mouse anti-Nav1.7 clone N68/6 monoclonal antibody used for behavioural experiments was developed by and obtained from the UC Davis/NIH NeuroMab Facility (Davis, CA, USA). The rabbit anti-guinea pig secondary biotin, streptavidin protein Dylight 488, and fluoromount G were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Cremophor, dimethyl sulphoxide (DMSO), naloxone HCl and sodium monoiodoacetate (MIA) were obtained from Sigma-Aldrich (Oakville, ON, Canada). Isoflurane and euthansol were purchased from CDMV (Dartmouth, NS, Canada).