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. 2017 Feb 2;8(26):42288–42299. doi: 10.18632/oncotarget.15039

Figure 4. WEE1 protects against double-stranded DNA breaks.

Figure 4

a. On the left, Western blot labelled with anti-pCDK1-Y15, anti-CDK1 and anti-β-Actin antibodies for CLMECs treated for 24 hr with vehicle control or 1 μM AZD1775. On the right, quantification of the pCDK1-Y15 band intensity divided by the CDK1 intensity (n=3 each). b. On the left, example flow cytometry data for CLMECs 24 hr after treatment with 1 μM AZD1775 or vehicle control. The vertical dotted lines separate different phases of the cell cycle. On the right, the mean percentage of cells in G0/G1, S and G2/M phases (n=3 each). AZD1775 data are compared with Controls for each phase. c. On the left, four example flow cytometry dot plots for unlabelled CLMECs (red) and CLMECs labelled with anti-γH2AX antibody (blue). The four conditions were vehicle control, 1 μM AZD1775, 1 μM AZD1775 + 10 μM RO-3306, and 1 μM AZD1775 + exogenous nucleosides (Nuc) (EmbryoMax®, 1:5 dilution) for 24 hr. On the right, mean data for the four groups (n=3 each). d. On the left, Western blot labelled with anti-pATM-S1981, anti-pCHK2-T68 and anti-β-Actin antibodies for CLMECs treated for 24 hr with vehicle control or 1 μM AZD1775. On the right, quantification of the pATM-S1981 and pCHK2-T68 band intensity divided by the β-Actin intensity (n=3 each). e. Mean data for the percentage fold increase in γH2AX positive LiECs and CLMECs with AZD1775 treatment (n=3 each).