Fitc conjugated anti mouse cd4 antibody

Recombinant murine IL-1β and IL-6 were purchased from PeproTech (Rocky Hill, NJ, USA). Recombinant human TGF-β1 was purchased from R&D systems (Minneapolis, MN, USA). PPARγ ligands (ciglitazone, rosiglitazone, and troglitazone) and trypan blue solution were obtained from Sigma- Aldrich (St. Louis, MO, USA). FITC-conjugated anti-mouse CD4 antibody, PE-conjugated anti-mouse IL-17 antibody, anti-mouse CD3 (145-2C11) antibody, and anti-mouse CD28 (37.51) antibody were purchased from BD Bioscience (Mississauga, ON, CA).

Mice with tumour formations were killed, and the tumours were collected. After straining through a 70-μm cell strainer (BD Biosciences), the tumour cells were washed two times and incubated with a PerCP-conjugated anti-mouse CD3e antibody (BD Biosciences), FITC-conjugated anti-mouse CD4 antibody (BD Biosciences), Alexa Fluor 647-conjugated anti-mouse CD8a antibody (BD Biosciences) or matched isotype controls. For analysis of MDSCs, cells were stained with PerCP-conjugated anti-mouse Gr-1 antibody (BD Biosciences) and APC-conjugated anti-mouse CD11b antibody (BD Biosciences) or matched isotype controls. Flow cytometric analysis was performed as described above.