Extended Data Fig. 2. Depletion of CD4 T cells and neutrophils.
(a) Flow cytometry gating strategy of tumour infiltrating leukocytes. (b) Normalized cell numbers of tumour infiltrating leukocytes after 14 days of tumour engraftment; from ZT1 to 21, n = 10, 9, 10, 7, 10, 8 mice from 4 independent experiments, Cosinor analysis. Treg, regulatory T cells. (c-d) Tumour volume upon anti-CD4 antibody depletion, n = 4 (ZT9 anti-CD4), n = 5 (ZT21 anti-CD4), n = 6 (control) mice (c), or anti Ly6G antibody depletion, n = 5 (control), n = 6 (anti-Ly6G) mice (d), from 2 independent experiments, two-way ANOVA. (e-j) Flow cytometry gating strategies and quantification of neutrophils in blood (e-h) or tumour (i-j) after anti-Ly6G treatment. Anti-mouse Ly6G antibody was given every 3 days, starting 1 day before the tumour inoculation (d-1). Neutrophil frequency after treatment at days 0 and 1 (e-f), from left to right, n = 3, 2, 3 mice, or day 12 (g-j), n = 3 (iso), n = 5 (anti-Ly6G) mice from 2 independent experiments, unpaired Student’s t-test. All data are represented as mean ± SEM, ns, not significant, all t-tests are two-tailed.