HeLa/RFP-H2B stable cell lines were seeded in an eight-chambered cover glass (Lab-Tek Chambered Borosilicate Coverglass System, Nunc). Cells were synchronized by T/T block and release following the procedure described above. Cells were placed in a 37°C-heated microscope chamber and observed under an inverted fluorescence microscope (Leica SP8) with a 63× lens. Images were acquired in the 561 channel every 5 min and captured for a total of 5 hours. Images were viewed and analyzed using Leica LAS AF Lite software. Laser micro-irradiation was performed as described previously (39). Briefly, cell cultured on glass-bottomed 35-mm dishes were micro-irradiated using a computer-controlled MicroPoint laser Ablation System (365 nm; Andor Company) coupled with an Olympus IX83 inverted microscope (60× oil-immersion objective). Time-lapse images of live cells were taken under the same microscope with the MetaMorph Microscope Automation & Image Analysis software.

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