Fig. 4.

Pulmonotoxic chemicals may induce DDR pathway activations dependent or independent of DNA strand breaks. a Fluorescence microscopy images showing the staining patterns of Hoechst and anti-γH2AX in single BEAS-2B cells treated with the indicated chemicals for 16 h. Cells with raw F₂ values (indicated below the cells) close to the average raw F₂ values across all the cells under the same treatment conditions are shown (scale bar = 30 µm). To allow visual comparisons, all the shown images have been scaled to the same intensity ranges with respect to the solvent controls. b Fluorescence microscopy images showing the DNA spots obtained from the Comet assays of BEAS-2B cells treated with the indicated chemicals for 4 h. DNA spots with % tail DNA values (indicated below the patterns) close to the average % tail DNA values across all the spots obtained under the same treatment conditions are shown (scale bar = 20 µm). To allow visual comparisons, all the shown images have been scaled to the same intensity ranges with respect to the solvent controls. c Changes in the median % tail DNA values obtained from the Comet assays of BEAS-2B cells treated with the indicated chemicals or solvent controls for 4 or 16 h (bars = mean of the log₂-ratio values obtained from at least three independent biological replicates, error bars = 95%-tile confidence intervals; X marks = ≤ 5% cells left). d Fluorescence microscopy images showing the staining patterns of Hoechst and anti-phospho-p53 (Ser15) in BEAS-2B cells treated with the indicated chemicals for 16 h. e Changes in the median nuclear phospho-p53 intensity levels of these cells quantified from the images with respect to the solvent controls (dots = mean of the median values, error bars = 95%-tile confidence intervals.) All P values shown in this figure are obtained from two-sided t tests and adjusted for false discovery rates (***P-adjusted ≤ 0.01, **P-adjusted ≤ 0.05, *P-adjusted ≤ 0.10)