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. 2015 Jul 22;6(29):28238–28256. doi: 10.18632/oncotarget.4958

Figure 8. vHMECs accumulate DNA damage with PDs and slowdown DNA replication.

Figure 8

A. Western blot analysis of protein extracts from control and treated vHMEC and vHMEC-hTERT cells. vHMEC and vHMEC-hTERT cells were exposed to 5Gy of gamma rays and processed 10 hours post-irradiation. vHMECs treated with 4 mM HU during 6 hours were used as positive control for collapsed replication forks. GAP120 was used as loading control. B. Cell cycle analysis of vHMEC cells at different PDs, as well as irradiated vHMECs and vHMECs-hTERT, was performed by measuring BrdU-Alexa488 levels and propidium iodide staining. BrdU-Alexa488 positive cells are colored in purple, dark green represents cells with 2N DNA content, pale green represents 4N DNA content cells, and the yellow dots are cells with 8N DNA content. Legend (upper right) shows the distribution of cells with different DNA content. C. Histograms showing intensity levels of BrdU-Alexa488 positive vHMECs throughout PDs as well as irradiated vHMECs. Dashed line indicates the cut-off for BrdU positive cells.