Fig. 1

Reduced smoker and enhanced LUAD donor BC proliferation. (A) BC counts over time in culture. LUAD cell numbers were significantly higher at day 10, and smoker cells lower at day 14. Data are presented as mean ± SEM (n ≥ 5; P < 0.05 for both by ANOVA). (B) Box and whisker plots showing differences in the fraction of EdU incorporating nuclei comparing control (never smoker and smoker) and cases (LUAD and LUSC) cells at day-7 (n ≥ 5, **P < 0.01 by T-test); (C) Representative immunofluorescent micrographs depicting cycling EdU-positive (green) and negative (DAPI-stained; blue) nuclei in BCs from the four groups. (D) Box and whisker plots showing percent of EdU incorporating nuclei from the studied groups in C (median and quartiles; n ≥ 5; *P < 0.05 and **P < 0.01 by ANOVA and T-test comparing controls and cases, respectively). (E) Representative flow cytometry dot-plots of cells treated with EdU for 1.5 h and counterstained with 7-AAD. Cells in the G0/G1-phase can be seen within the bottom left box; S-phase cells in the top elongated box, and G2/M in the bottom right box. (F) Median and quartiles of cells in the S-phase of the cell cycle from panel E by group (n ≥ 5; *P < 0.05 and **P < 0.01 by ANOVA). Passage 2 never-smoker (Never) and smoker controls, and lung adenocarcinoma (LUAD), and squamous cell carcinoma (LUSC) cells were used in these experiments. Scale bar = 20 μm