Figure 4.

Effects of neutrophil depletion on Tregs. ID8-KRAS-injected mice were treated with anti-Ly6G mAb (n=9) or isotype control mAb (n=8) from day 10 after the inoculation of ID8-KRAS cells and sacrificed on day 16. No cancer mice were used as controls (n=6). Ascites and spleen were obtained from the no cancer mice and ID8-KRAS mice treated with anti-Ly6G or isotype mAb after washing with 3 ml of PBS. T cells were collected by Percoll density gradient (44%/70%). T cells were stained with the following antibodies: FITC-anti-mouse CD25, APC-anti-mouse CD4, and PE-anti-mouse FoxP3. The FoxP3⁺CD25⁺/CD4⁺ ratio of (A) ascites and (B) spleen was analyzed by flow cytometry. Error bars represent the means ± SEM. Statistical analysis was performed using the Student's t-test. The P-value was adjusted using the Holm's method (^*P<0.05).