Hrp labeled goat anti mouse igg h l

Drug FGF401 was purchased from CASYMCHEM; BLU-554, Geldanamycin, Imatinib, LY-294002, Parthenolide, Tanespimecin, Trichostatin A and Vorinostat were purchased from MedChemExpress. Drug details have shown in Supplementary Table S2. Antibodies FGFR4, p-ERK 1/2 and ERK 1/2 were purchased from Abclonal; Antibodies FGFR4, p-AKT (T308) and AKT were purchased from Cell Signaling Technology; Antibodies FGF19 and KLB were purchased from Abcam. HRP labeled goat anti rabbit IgG (H + L) and HRP labeled goat anti mouse IgG (H + L) were purchased from Beyotime. Antibodies details have shown in Supplementary Table S3.

Cordycepin (>98% purity, C₁₀H₁₃N₅O₃) was provided by Prof. Hang-Hai Li, South China Normal University, China. Phentolamine was purchased from Shanghai Xudong Haipu Pharmaceutical company (China) and sodium nitroprusside was purchased from the Guangdong Hongyuan Group Pharmaceutical company (China). Other major chemicals used in the experiments included the antagonist of A1R, DPCPX (Tocris Company, Cat#0439), and the antagonist of A2AR, SCH58261 (Tocris Company, Cat# 2270), anti-Adenosine A1 receptor antibody (Abcam, Cat# ab82477), anti-Adenosine receptor A2A antibody (Abcam, Cat# ab3461), monoclonal Anti-γ-Tubulin antibody (Sigma-Aldrich, Cat# T6557), HRP-labeled Goat Anti-Rabbit IgG (H+L) (Beyotime biotechnology, Cat# A0208), and HRP-labeled Goat Anti-Mouse IgG (H+L) (Beyotime biotechnology, Cat# A0216).

Sheep (ALC and MF) and mouse skin tissues were fixed in 4% paraformaldehyde formalin in PBS at 4 °C overnight, embedded in paraffin, sectioned at 5 μm, and stained with a hematoxylin and eosin staining kit (H&E). The following antibodies were used for immunostaining: anti-Ki67 (Abcam, ab15580), anti-Caspase 3 (Abcam, ab184787), anti-AKT1 + AKT2 + AKT3 (Abcam, ab179463); phospho-PI3-kinase p85-alpha/gamma (Abmart, T40116F); mTOR antibody (Abmart, T55306); HRP-labeled goat anti-rabbit IgG(H+L) (Beyotime, A0208), and HRP-labeled goat anti-mouse IgG(H+L) (Beyotime, A0216). The signal was detected using the DAB Horseradish Peroxidase Color Development Kit (Beyotime, P0202), and the sections were stained with hematoxylin. Photographs were taken using a microscope (ECHO, RVL-100-G, USA).

Sheep polyclonal anti-Corticotropin Releasing Factor Antibody (NB110-81721, Novus, USA); rabbit anti-c-Fos mAb (2250s, Cell Signaling Technology, MA, USA); γ-aminobutyric acid (A2129-25G, Sigma-Aldrich LC, USA); muscimol hydrobromide (G019, Sigma-Aldrich); mouse monoclonal Anti-GAD 67 (MAB5406, Millipore, USA); rabbit polyclonal anti-GAD 67 (10408-1-AP, Proteintech, USA); mouse polyclonal anti-GAD 65 (20746-1-AP, Proteintech, USA); mouse anti-β-actin mAb (sc-47778, Santa Cruz, CA, USA); rabbit anti-GAPDH pAb (AC001, Abclonal, MA, USA); Alexa Fluor 488 donkey anti-Rabbit IgG (H + L) and Alexa Fluor 594 donkey anti-Sheep IgG (H + L) (Life Technologies, CA, USA); DAPI staining solution (C1005, Beyotime, China); alkaline phosphatase horse anti-mouse IgG (ZB-2310, Zsbio, China); HRP-labeled goat anti-mouse IgG (H + L) (A0216, Beyotime, China); BCA protein assay kit (P0012 Beyotime, China); RIPA lysis buffer (P00138, Beyotime, China); phenyl-methanesulfonyl fluoride (PMSF) (ST506, Beyotime, China); sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) sample loading buffer (P0015, Beyotime, China); BCIP/NBT alkaline phosphatase color development kit (C3206, Beyotime, China); BeyoECL Moon kit (P0018, Beyotime, China).

Mouse ears or cultured DCs were lysed in radio immunoprecipitation assay buffer supplemented with protease and phosphatase inhibitor cocktail (Thermo Scientific, #78440). Mouse anti‐β‐actin antibody (Ab) (1:2,000; Cell Signaling Technology, #3700S), rabbit anti‐RIG‐I Ab (1:1,000; Abcam, #ab45428), rabbit anti‐p‐IκBα Ab (1:1,000; Cell Signaling Technology, #2859), HRP‐labeled goat anti‐mouse IgG(H+L) (1:2,000; Beyotime, #A0216), and HRP‐labeled goat anti‐rabbit IgG(H+L) (1:1,000; Beyotime, #A0208) were used. The signal was detected with ECL Western Blotting Substrate (Thermo Scientific, #34095) and GE ImageQuant LAS 4000 (GE Healthcare) or Amersham Imager 600 (GE Healthcare). Images have been cropped for presentation.