Ab180780

RPMI 1640 media, Fetal Bovine Serum (FBS) and l-glutamine, phosphate buffer saline (PBS) were purchased from HiMedia (Mumbai, India). Penicillin and streptomycin were obtained from Life Technologies, USA. Antibodies against IL-2 (ab180780), IL-6 (ab208113), anti-mouse HRP linked secondary antibody, anti-rabbit HRP linked secondary antibody were purchased from Abcam (MA, USA). Antibodies against TGF-β, Actin were purchased from Santa Cruz Biotechnology (Dallas, Texas USA). Anti-CD4, anti-CD8, anti-CD19, anti CD3e antibody (FITC conjugated), anti Ki67 antibody (PE conjugated), anti-IL-2, and IL-6 antibodies (PE conjugated) were purchased from BD Biosciences, USA. PMA, Ionomycin, Brefeldin A, saponin were purchased from Sigma Aldrich, India. Vectashield mounting medium was from Vector Laboratories, Inc. (CA, USA). MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide), Bovine serum albumin (BSA) were purchased from SRL India. SRBC was procured from Indian Veterinary Research Institute, Eastern Regional Station, Kolkata. Cyclophosphamide was purchased from Abcam.

Immunostaining of the formalin-fixed, paraffin-embedded mice spleen tissue from each group was started with overnight incubation with primary antibodies against IL-2 (ab180780; Abcam) and IL-6 (ab208113; Abcam) and was completed according to the protocol (MP-7601; IHC Guide, Vector Laboratories). Bright field images were captured with a microscope (Leica).