Fig. 8

NP-cGAMP inhalation activates CD8 T cells and improves the ratio of CD8⁺ T/Treg in 4T1 lung metastases. Twenty four hours after the last inhalation, the mice under indicated treatment (n = 6 biologically independent mice/group) were sacrificed and both metastases-bearing lungs were dissected for FACS and immunohistochemical studies. a After ex vivo re-stimulation of T cells with phorbol 12-myristate 13-acetate/ionomycin, representative FACS dot plots (n = 5000) of activated CD8⁺ T cells by staining intracellular IFNγ from the left and right (IR) lung of each indicated treatment. b Quantitative analysis of frequency of IFN-γ⁺CD8⁺ T cells in the left and right lung. c Representative FACS analysis and d quantitative frequency of FoxP3⁺CD4⁺ Treg cells and ratio of CD8/Treg. e Immunohistochemical staining of CD8⁺ T cells and FoxP3⁺ Tregs in metastases-bearing lung tissues of n = 3 biologically independent mice under indicated treatment, and quantitative data showing the mean ± SD ratio of CD8/Treg in lung metastases. f NP-CTR: NP-2′5′-GpAp as a negative control of NP-cGAMP; IR (L) and IR (R): left (non-irradiated) and right (irradiated) lung of IR alone; the same applies to NP-cGAMP + IR(L) or IR(R). Scale bar = 20 μm. *P < 0.05; ***p < 0.001 by Student’s T test. Source data are provided as a Source Data file