Fig. 4

Cell cycle profile of hMECs with IR treatment and time. a Immunofluorescence staining of hMECs with DAPI (blue) for DNA content and the nucleoside analogue EdU (green) for newly synthesised DNA. b Representative distribution of total DAPI intensity for hMECs at baseline (0 Gy and 0 hours). The cell cycle phases are labelled, where the first peak represents cells with 2n DNA content (G₁) and the second peak represents cells with 4n DNA content (G₂/M). The distribution of total DAPI intensity is modelled using t distributions (solid red lines), and the intersection of these distributions (dashed red lines) are used to assign cell cycle phases. c Scatter plot of total EdU intensity against total DAPI intensity on a cell-by-cell basis. The relationship between the cell cycle phases (G₁, S and G₂/M) and total DAPI intensity (DNA content) vs. total EdU intensity (newly synthesised DNA) is indicated. d Mean proportion of cells within each cell cycle phase for each hMEC subpopulation at 0 Gy (± SEM). Cell cycle phase proportions from donor Q687. e Cell cycle phase distributions of hMECs post 5 Gy IR with time. Distributions of total DAPI intensity for all hMECs (first column) and stratified per hMEC subpopulation (second column) from donor Q685 post 5 Gy IR over 48 hours. DAPI 4′,6-diamidino-2-phenylindole