Fig. 7: Increased phagocytic activity of macrophages mediated by cytotoxic CD8+ T-cell-dependent killing of cancer cells in PLD1 inhibition.

a MC38-GFP-Luc cells were orthotopically injected into the cecal submucosa of mice and then treated with A3373 (10 mg/kg) every day for 4 weeks. The mortality of tumor-bearing mice was analyzed using Kaplan–Meier analysis. b The tumors that developed at the injection site were photographed by an optical camera, and their weight was quantified. c IHC of active caspase-3 and Ki67 in the tumor tissues. Scale bar, 20 µm. d The levels of “do not eat-me” signals in the cancer cells isolated from vehicle- or A3373-treated tumor-bearing mice were analyzed by flow cytometry. e IHC of CRT in the tumor tissues. Scale bar, 20 µm. f The MC38-GFP-Luc cells derived from A3373-treated mice were cocultured with BMDMs for 2 h, and phagocytosis activity was analyzed by flow cytometry. The population of GFP⁺CD11b⁺ macrophages was quantified. g Cytotoxic T-cell-mediated killing assay of MC38-GFP-Luc cells derived from orthotopically injected tumors. h The populations of CD107a⁺ granzyme B⁺ and FasL⁺IFNγ⁺ in activated CD8⁺ T cells were analyzed by flow cytometry. i IFNγ⁺, IL17a⁺, and Foxp3⁺ cells in activated CD4⁺ T cells were analyzed by flow cytometry. The results are representative of at least three independent experiments and presented as the means ± SDs. **P < 0.01; ***P < 0.001, n.s. not significant.