Fig. 3. KACs evolve early and before tumour onset during tobacco-associated KM-LUAD pathogenesis.
a, Schematic view of the in vivo experimental design. b, Fraction of malignant cells (left) and KACs (right) across treatment groups and time points. Box-and-whisker definitions are same as in Fig. 1g. n = 4 biologically independent samples per condition. P values were calculated using two-sided Mann–Whitney U-test. NS, not significant. c, IF analysis of KRT8, LAMP3 and PDPN in mouse lung tissues. Scale bar, 10 μm. Results are representative of two independent biological replicates per treatment and timepoint. Staining was repeated three times with similar results. d, Top, distribution of CNV scores among alveolar and malignant cells. n on top of each bar denotes the numbers of KrasG12D mutant cells in each cell group. Bottom, fraction of KrasG12D mutant cells in KACs, malignant, AT1 and AT2 subsets. n = 496 (AT1), 1,320 (AT2), 512 (KACs) and 1,503 (malignant) cells. P values were calculated using two-sided Mann–Whitney U-test with Benjamini–Hochberg correction. e, ST analysis of lung tissue at 7 months after exposure to NNK and showing histological annotation of H&E-stained Visium slide (left) and spatial heatmaps showing scaled expression of KRT8 as well as KAC and KRAS signatures. ST analysis was done on three different tumour-bearing mouse lung tissues from two mice at 7 months following NNK. The schematic in a was created using BioRender (https://www.biorender.com).