α galactosylceramide α galcer

Anti-mouse CD4- and CD8-coated magnetic beads were obtained from Miltenyi Biotech (Bergisch Gladbach, Germany). C57BL/6 mice were purchased from Joint Ventures Sipper BK Experimental Animal Co. (Shanghai, China) RPMI medium 1640 and FBS were purchased from PAA Laboratories (Freiburg, Germany). PE-CY7-conjugated anti-mouse CD3, APC-conjugated anti-mouse CD8, FITC-conjugated anti-mouse CD4 and PE-conjugated anti-mouse IFN-γ antibodies were purchased from eBioscience. PE-conjugated anti-mouse CD24 antibodies were purchased from Acris Antibodies (Rockville, MD, USA). Antibodies specific to p-ERK (Thr202/Tyr204), p-JNK (Thr183/Tyr185), p-p38 (Thr180/Tyr182), p-p65 (Ser536), p-PLCγ1 (Tyr783), p-Zap70 (Tyr493), p-LCK (Tyr505), LCK and β-actin were obtained from Cell Signaling Technology (Danvers, MA, USA). Trizol reagents were from Invitrogen (Carlsbad, CA, USA). Prime Script RT-PCR Kit and SYBR Premix ExTaq kit were from Takara Bio. Concanavalin A (ConA) was obtained from Sigma-Aldrich (Saint Louis, MO, USA), and α-galactosylceramide (α-GalCer) was obtained from Enzo Life Sciences (Farmingdale, NY, USA). A monoclonal antibody specific to CD4 (GK1.5) was obtained from BioXCell (West Lebanon, NH, USA).