Gm csf

Proteins were extracted from the spleens using lysis buffer (Applygen) plus protease inhibitors and phosphatase inhibitors (Roche), then subjected to SDS-PAGE followed by transfer to nitrocellulose membranes (Pall) and probing with GM-CSF (Boster) and GAPDH primary antibodies (Santa Cruz). The specific bands were visualized with an ECL Detection Kit (Pierce), and images were acquired using Kodak X-Omat film.

Dulbecco’s Modified Eagle Medium (12100046), RPMI-1640 (31800022), L-glutamine (A2916801), penicillin G (15140-122), streptomycin (15140-122), amphotericin B (15290-017), and all other cell-culture reagents were purchased from Gibco BRL Life Technologies (Grand Island, NY, USA). Concanavalin A (con A) (L7647) and dimethyl sulfoxide (DMSO) (D2650) were obtained from the Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). The Multiplex ELISA Kit For Mouse Cytokine (MEK1006, GMCSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, IL-12p70, IL-17, MCP-1, MIP-1α, RANTES, TNFα) was obtained from Boster Biological Technology (Pleasanton, CA, USA). Certified fetal bovine serum (FBS) (04-001-1A) was obtained from Biological Industries (Kibbutz Beit Haemek, Israel).