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. 2017 Jun 7;7:2927. doi: 10.1038/s41598-017-03060-w

Figure 5.

Figure 5

Cdc6 is important for E7 expressing cells to alleviate G1 arrest under hypoxia. (a) Cdc6 in RPE1 E7 cells was knocked down by siRNA at indicated doses and analyzed by immunoblotting (Upper panel) (original blots are presented in Supplementary Figure 3) and RT-qPCR (Lower panel, 3 nM siRNA was used). (b) RPE1 E7 cells were transfected with either Cdc6 or negative control siRNA. Cells were treated with 200 μM DFO 24 hours post-transfection and cultured for an additional 8 hours, stained with PI and analyzed by flow cytometry. A representative experiment of 3 is shown (Upper panel) and summarized (Lower panel). (c) RPE1 E7 cells were transfected with either Cdc6 or negative control siRNA. Twenty-four hours later, cells were treated with 200 μM DFO for 6 hours and then labeled with BrdU for 2 additional hours. Cells were stained with anti-BrdU antibody, counterstained with 7-AAD, analyzed by flow cytometry. Data from a representative of 3 experiments are shown (Upper panel). Data are summarized (Lower panel). Error bars reflected the standard error of the mean. *P < 0.05; **P < 0.01.