Mice were sacrificed 20 hours after induction of CASP by cervival dislocation under deep anaesthesia. Spleens were taken and splenocytes were isolated. After blocking with FcBlock (BD Biosciences, San Jose, California, USA) cells were stained with appropriate antibodies according to the manufacturer’s instructions. PE-conjugated anti-mouse DR5 and PE-conjugated anti-mouse TRAIL were purchased from eBioscience. Isotype controls (PE-conjugated Armenian Hamster IgG and Rat IgG2ak) were also purchased from eBioscience. APC- and FITC-conjugated anti-Ly6G (clone 1A8) was purchased from Miltenyi (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany). Rat IgG2a (Miltenyi) was used for isotype control. FITC-conjugated anti-B220, FITC-conjugated anti-CD3, PE-conjugated anti-CD11b and FITC-conjugated anti-Ly6C was purchased from Pharmingen (BD Biosciences, San Jose, California, USA). Appropriate isotype controls were also purchased from Pharmingen.
FACS analysis was done using a BD FACS Calibur system. For cell analysis FlowJo (Treestar Inc., Ashland, USA) and WinMDI software were used.
FACS analysis was done using a BD FACS Calibur system. For cell analysis FlowJo (Treestar Inc., Ashland, USA) and WinMDI software were used.