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. 2015 Dec 23;14(24):3864–3876. doi: 10.1080/15384101.2015.1120914

Figure 2.

Figure 2.

Thymidine increases DNA damage in Erk5-depleted Jurkat cells. (A) Viability of cells after 18 h treatment with 5 μg/mL aphidicolin or 2 mM HU. (Left) Annexin V/DAPI analysis of cells treated with aphidicolin or HU. The numbers inside the plot indicate the percentage of annexin V-positive cells in this particular experiment. (Right) Quantification of apoptosis, n=3, *p<0.05. (B) Representative western blot showing phospho-H2AX (p-H2AX) in shCtrl and shErk5 cell lines at the indicated times after release from an 18 h thymidine block. Histone 3 was used as loading control. (Right) Densitometric analysis of the protein gel blot, the ratio phospho-H2AX:H3 ± s.d. is shown, n=3. (C) Representative cell cycle profiles after 0 h, 8 h, 16 h or 24 h release from 18 h thymidine block, as analyzed by PI staining and flow cytometry. (Right) Quantification of subG1 fraction after release from thymidine block, n=6. A pool of 2 shCtrl or 2 shErk5 cell lines was used in these experiments (A-C). Paired Student's t-test, *p<0.05 **p<0.01.