Dexamethasone does not completely suppress tumor-specific T cell activities in antigen-naïve mice.

Healthy mice received activated pmel-1 T cells labeled with the congenic marker Thy1.1 on day −2, followed by daily administration of dexamethasone (3 mg/kg) from day −2 to day 6. On day 0, mice were vaccinated with gp100, and spleens were harvested for flow cytometry analysis on day 7. The cells were stained with Zombie Yellow to eliminate dead cells. The cells were then pretreated with Fc Block, stained with FITC-conjugated anti-Thy1.1, PerCP/Cyanine5.5-conjugated anti-CD4, APC-conjugated anti-CD3, APC/Cyanine7-conjugated anti-CD8, PE/Dazzle594-conjugated anti-B220, anti-F4/80, anti-NK1.1 and anti-Ly6G, followed by fixation, permeabilization and staining with PE-conjugated anti-Foxp3 and PE/Cyanine7-conjugated anti-Ki67 (Thermo Fisher Scientific) using Foxp3/Transcription Factor Staining Buffer Set (Thermo Fisher Scientific) according to the manufacturer's protocols.