Ad5-TRAIL/CpG therapy alters MDSC location within the tumor-bearing kidney. BALB/c mice were implanted IR with 2 × 105 Renca cells and treated with PBS or Ad5-TRAIL/CpG on day 7 (some mice received no injection). Tumor-bearing and contralateral kidneys were harvested on day 12. Prior to harvest, all mice were injected i.v. with PE-conjugated anti-CD45.2 mAb (3 μg) in PBS to discriminate tissue- from vasculature-associated MDSC, and killed 3 min later. Single-cell suspensions of tumor-bearing kidneys were prepared and stained for flow cytometric analyses to assess MDSC cell infiltration. a Representative flow plots for tissue- or vasculature-localized MDSC in tumor-bearing and contralateral kidneys from untreated mice. b, c Mean (±SEM) number and frequency of tissue- and vasculature-localized MDSC from (b) tumor-bearing or (c) contralateral kidneys. Data are representative of three independent experiments totaling 8 mice. d, e Mean (±SEM) number and frequency of bulk MDSC, Ly6G+, or Ly6G− MDSC in the tissue (d) and vasculature (e) of tumor-bearing kidneys from mice treated with either PBS or Ad5-TRAIL/CpG on day 7. Data are representative of four independent experiments with at least 4 mice/group/experiment. For all experiments **p ≤ 0.01, ***p ≤ 0.001, n.s. not significant using Student’s t test