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. 2017 Jun 12;36(14):2161–2176. doi: 10.15252/embj.201696082

Figure 5. ATM activity is detected throughout chromatin upon localized DNA damage.

Figure 5

  1. H2B‐ATKAR 1/FRET spreads over chromatin after localized damage. U2OS cells expressing H2B‐ATKAR were laser‐microirradiated, and 1/FRET was followed over time. Note that bleaching of fluorophores precludes FRET analysis within the laser‐microirradiated area.
  2. Quantification of spread of H2B‐ATKAR (wt) or non‐phosphorylatable Ala‐H2B‐ATKAR (Ala) FRET change after laser microirradiation in U2OS cells. Measurements were performed distal to the laser‐microirradiated area. Graph shows average and SD of at least seven cells.
  3. H2B‐ATKAR detects both ATM and ATR activity after laser microirradiation. Quantification of spread of H2B‐ATKAR FRET change after laser microirradiation in U2OS cells in the presence of ATM or ATR inhibitors. Inhibitors were added 30 min before laser microirradiation. Graph shows average and SD of at least five cells. Measurements were performed as in (B).