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. 2018 Mar 29;19(5):e44722. doi: 10.15252/embr.201744722

Figure 1. Repeated exposure to arsenite decreases the number of cells which commit to the senescence process.

Figure 1

  1. (left) IDH4 cells were treated daily post‐induction of senescence for 30 min with (AS) or without (UNT) 0.5 mM sodium arsenite. Proliferating (PRO, Days 0–3), presenescent (PRE, days 4–6) and senescent (SEN, days 7–10) cells were subsequently subjected to staining for β‐galactosidase activity. Phase contrast images demonstrate the β‐galactosidase staining of the IDH4 cells at the various stages of the senescence process. Scale bars, 400 μm. (right) Graph represents the percentage of cells that stained positive for β‐galactosidase activity (stained blue‐green) in the phase contrast images shown in (left panel). The percentage of senescent cells in each experiment was calculated using three random fields. Data are represented as a mean of three independent experiments ± SE (error bars). *P < 0.05, **P < 0.01 (Student's t‐test).
  2. Whole‐cell extracts from IDH4 cells treated (AS) or not (UNT) with sodium arsenite as indicated above were prepared and analyzed by Western blot using antibodies for p53, p21, p16, and tubulin (loading control).
  3. Whole‐cell extracts from IDH4 cells treated (AS) or not (UNT) with sodium arsenite as indicated above were prepared and analyzed by Western blot using antibodies that detect caspase‐3 cleavage products and tubulin (loading control). HeLa cells treated with 1 μM staurosporine for 3 h were used as a positive control for caspase‐3 cleavage (lane 7).

Source data are available online for this figure.