Mc903

MC903 (Calcipotriol; R and D Systems) was applied to the shaved mouse cheek (20 μl of 0.2 mM in ethanol) or rostral back (40 µl of 0.2 mM in ethanol) once per day for 1–12 days using a pipette. 100% ethanol was used. All MC903 studies were performed on 8–12 week old age-matched mice. Behavior, RNA-seq, flow cytometry, and immunohistochemistry were performed on days 1, 2, 3, 5, eight and/or 12. For AMG 487 experiments in the MC903 model, 50 µL 3.31 mM AMG 487 (Tocris) or 20% HPCD-PBS vehicle was injected subcutaneously one hour prior to recording behavior (Qu et al., 2015 (link)). Spontaneous scratching was manually scored for the first 30 min of observation. Both bout number and length were recorded. Behavioral scoring was performed while blind to experimental condition and mouse genotype.

Agmatine sulfate salt (Cat#a7127), histamine (Cat#y0001779), compound 48/80 (Cat#C2313), resiniferatoxin (RTX, Cat#R8756), chlorpheniramine maleate (Cat#C3025), chloroquine (CQ, Cat#C6628), naloxone hydrochloride (Cat#N3136), amiloride hydrochloride (Cat#1019701) were all obtained from Sigma-Aldrich (St. Louis, MO, United States). MC903 (Cat# 2700/10) was purchased from R&D systems (Minneapolis, MN, United States). Morphine hydrochloride was obtained from China Northeast Pharmaceutical Group Shenyang No. 1 Pharmaceutical Co., Ltd (Shenyang City, China). naloxone hydrochloride HC-030031 (Cat#2896), capsazepine (CPZ, Cat#0464) and APETx2 (Cat#4804) were purchased from Tocris (Minneapolis, MN, United States). ZnCl₂ was obtained from National Pharmaceutical Group pharmaceutical Co., Ltd (Beijing, China). RTX and HC-030031 were freshly dissolved in 10% DMSO. Other reagents were dissolved in saline. Information relating to timing and doses are indicated in the results section or figure legends.