Fixation and permeabilization solution kit

Flow cytometry protocol has previously described^{61 (link),62 (link)}. The single-cell suspensions of tumor-burdened lungs, bronchial dLN or the obtained LILs were re-suspended in FACS buffer (PBS, 1%BSA) and blocked with anti-mouse CD16/32 antibodies for 10 min prior to staining with the antibodies of the surface markers. For intracellular cytokine staining, cells were fixed and permealized with a fixation and permeabilization solution kit (Cat# 424401, Biolegend) followed by staining with the specific antibodies against intracellular cytokines. Antibodies used for flow cytometry analysis were listed in Table S5. Flow cytometry data were acquired on a FACSCelesta flow cytometer (BD Biosciences, BD FACSDiVa Software v8.0.1.1) and analyzed with Flowjo 10.6.2 software (TreeStar). The staining antibodies were listed in Supplementary Table 7.

Flow cytometry protocol has been previously described.^[43] The single‐cell suspensions of tumor‐burdened lungs, bronchial dLN or the obtained LILs were re‐suspended in FACS buffer (PBS, 1% BSA) and blocked with anti‐mouse CD16/32 antibodies for 10 min prior to staining with the antibodies of the surface markers. For intracellular cytokine staining, cells were fixed and permealized with a fixation and permeabilization solution kit (Cat# 424 401, Biolegend) followed by staining with the specific antibodies against intracellular cytokines. Antibodies used for flow cytometry analysis were listed in Table S9, SUpporting Information. Flow cytometry data were acquired on a FACSCelesta or LSRFortessaX20 flow cytometer (BD Biosciences) and analyzed with FlowJo X software (TreeStar).