Extended Data Fig. 4: Immune cell profiling in response to PM.

A) Immune cell frequencies in the lungs determined by flow cytometry 24 h post-exposure from induced T and ET mice after 50 μg PM (red) or PBS control (blue) (n = 8 mice per group). Data are presented as the frequency among live CD45+ immune cells. One-way ANOVA. B) Representative immunofluorescent images of CD68+ macrophages (cyan) and tdTomato+ EGFR mutant cells (red) within ET lungs exposed to control or 50 μg PM. Quantification of CD68+ cells per mm² of lung tissue (n = 4 mice per group). One-way ANOVA. C) Representative immunofluorescent images of CD68 (red), CD11b (green) and merged images from induced ET mice after 3 weeks of exposure to PBS (top) or 50 μg PM (bottom). Quantification of alveolar macrophages (AMΦ, CD68+CD11b−) and interstitial macrophages (IMΦ, CD68+CD11b+) per mm² of lung tissue, selecting 10 x random 500 μm² fields of view per mouse (n = 3 mice per group). One-way ANOVA. D) Representative immunofluorescent images of CD68+ macrophages (cyan) within CCSP-rtTA; TetO-EGFR^L858R lungs treated with PBS (top) or 50 μg PM (bottom) 10 weeks post oncogene induction; quantification of CD68+ cells per mm² of lung tissue, selecting 20 x random 500 μm² fields of view per mouse (n = 3 mice per group). Unpaired t-test. E) Representative immunofluorescent images of CD68+ macrophages (cyan) and tdTomato+ Kras^G12D mutant cells (red) within KT lungs treated with PBS (top panel) or 50 μg PM (bottom) 10 weeks post oncogene induction; quantification of CD68+ cells per mm² of lung tissue, selecting 20 x 500 μm² fields of view containing RFP+ cells per mouse (n = 3 mice per group). Unpaired t-test. Scale bar 50 µm B & D, 150 µm C & E. Gating strategies for flow cytometry analysis provided in Extended Data Fig. 6.

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