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. 2020 Feb 25;21(4):e49076. doi: 10.15252/embr.201949076

Figure 4. Telomere fusions in HeLa cells after RAP1 depletion.

Figure 4

  1. RAP1 expression in HeLa cells treated with the telomerase inhibitor BIBR1532 for 25 days (20 μM final concentration). Fifteen days before cell harvesting, doxycycline (DOX; 1 μg/μl final concentration) was added to deplete the expression of RAP1.
  2. Telomere length analysis by Southern blotting of the samples described in (A). The size of the main intensity peak is indicated at the bottom of the gel.
  3. Number of fusions in HeLa cells after 25 days in culture. Cells were maintained with BIBR1532 during the whole period of the experiment, while doxycycline (1 μg/μl final concentration) and the infection with shControl, shLIG3, or shLIG4 were carried out for the last 15 days of the experiment. Data represent mean ± SD of three biological replicates. Statistical analyses were performed using Mann–Whitney U‐test (*P < 0.05; **P < 0.001; ***P < 0.0001).
  4. Examples of metaphase spreads hybridized with a telomeric PNA probe (green) when RAP1 was depleted (+DOX). Telomerase was inhibited with BIBR1532 for 25 days. Scale bar = 10 μm.
  5. Quantification of chromosome aberrations observed in RAP1‐depleted cells (+DOX) or control (‐DOX). Data represent mean ± SD of three biological replicates. Approximately 15 metaphase spreads were analyzed per replicate with a total of 2,300 chromosomes examined per condition. *P < 0.05; two‐tailed Student's t‐test.