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. 2015 Jun 25;11(6):e1005167. doi: 10.1371/journal.pgen.1005167

Fig 7. AKTIP downregulation impairs telomere replication.

Fig 7

Ctr or shAKTIP HeLa cells were synchronized at the G1/S boundary with a double thymidine block and harvested at the indicated times. (A) Scatter plots showing the proportions of cells in S phase in asynchronous cultures (As) and in cultures analyzed at various times after release from the double thymidine block. Prior to harvest at each time point, the cells were incubated with BrdU for 30 min. (B) ChIP analysis on synchronized HeLa cells incubated with BrdU for 1 h before harvesting. Precipitations were performed with an anti-TRF1 antibody. IgG antibody was used as negative control. Inputs represent 10 and 1% of genomic DNA. Dot-blot analysis was performed using telomeric or ALU repeat-specific probes. Precipitated DNA was analyzed by Western blotting with an anti-BrdU antibody. (C, D) Quantification of the data expressed in arbitrary units (A.U.) of unlabeled (C) or BrdU-labeled (D) precipitated telomeric DNA at the different time points of analysis, each normalized to input samples. The graphs show three independent experiments, with error bars indicating the SD.