Anti n cadherin

Total proteins were extracted from transfected cells using RIPA lysis buffer containing PMSF (Solarbio) and protease inhibitor cocktail (Promega) according to the instructions of the manufacturer. The protein concentration was determined by BCA Protein Assay Kit (Multi Sciences). After mixed with loading buffer and heated at 99°C for 5 minutes, the protein lysates were separated by 10% sodium dodecyl sulphate‐polyacrylamide gel electrophoresis and transferred onto PVDF membranes (Millipore). The transferred membranes were blocked with 5% skim milk for 1 hour at room temperature and incubated overnight with the specific primary antibodies at 4°C. Subsequently, the membranes were incubated at room temperature with horseradish peroxidase–conjugated goat anti‐rabbit IgG (KPL) for 1 hour. The bands were visualized with enhanced chemiluminescence (ECL) detection reagent (Multi Sciences) by Chemi XT 4 (Syngene). The primary antibodies were used: anti‐β‐actin (AC026, ABclonal), anti‐E‐cadherin (E‐AB‐35932, Elabscience), anti‐N‐cadherin (E‐AB‐64011, Elabscience) and anti‐FAM83H (ab121816, Abcam).

Dulbecco’s modified Eagle’s medium (DMEM), bovine serum albumin (BSA), dithiothreitol, phenylmethylsulfonylfluoride (PMSF) were obtained from Sigma Chemical, Co. (St. Louis, MO, United States); VD3 was obtained from DBA Italia (Segrate, Milan, Italy); anti-GFAP antibody was obtained from Dako, Agilent (Santa Clara, CA, United States), anti-N-cadherin, anti- peroxisome proliferator-activated receptor gamma (PPARγ), anti-VDR from Elabscience (Houston, TX, United States) and anti-beta actin antibodies were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, United States); anti-neuron specific enolase (NSE) and anti-NF200 antibodies were from NOVOCASTRA Laboratories, Ltd. (Newcastle, United Kingdom). For research involving biohazards, biological select agents and reagents, standard biosecurity safety procedures were carried out.