Fig. 2.

Phagocytes are crucial to antitumour effect of ROCK blockade. a–c CT26.CL25 tumour-bearing BALB/c mice (a) and B16F10-Ova tumour-bearing C57BL/6 mice (b) were injected (i.v.) with Y27632 (10 mg kg⁻¹) or vehicle (PBS). Tumour dimensions were measured at the indicated times using calipers and tumour volume was calculated (mm³). Data are presented as means ± s.e.m. (left panels) and tumour growth curves of individual mice (right panels) (CT26.CL25 model, n = 17; B16F10-Ova model, n = 17–19). c Tumour weights in individual mice were analysed at the end of experiments. Each dot represents tumour weight of individual mice. Bars indicate mean values. d–g CT26.CL25-mCherry tumour-bearing BALB/c mice were injected (i.v.) with Y27632 (10 mg kg⁻¹) together with clodronate liposomes or PBS liposomes as described in Supplementary Fig. 6c. d Tumour volume was measured at the indicated time. Data are presented as means ± s.e.m. (left panel) and tumour growth curves of individual mice (right panels) (n = 9). e, f For the PBS liposomes-treated group, F4/80⁺ macrophages or CD11c⁺ DCs were isolated from tumours, and the percentage of cells harbouring mCherry⁺ tumour cells was determined by flow cytometry. e Representative flow cytometry plots. The grey peaks represent mCherry levels in macrophages and DCs from CT26.CL25 tumour-bearing mice (mCherry-negative tumour). f Data are presented as means ± s.d. (n = 3). g At 22 days after tumour inoculation, single-cell suspensions from tumour-draining LNs were stimulated with β-gal-derived peptide for 48 h, and secreted IFN-γ was measured by ELISA. Data are presented as means ± s.d. (n = 4). h At 22 days after tumour inoculation, CD8⁺ and CD8⁻ cells were isolated from tumour-draining LNs and stimulated with β-gal-derived peptide for 48 h, and secreted IFN-γ was measured by ELISA. Data are presented as means ± s.d. (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001; significance determined by unpaired Student’s t test (a, f), Mann–Whitney test (b, c) or one-way ANOVA with Tukey’s post hoc test (d, g, h)