Figure 2.
Figure 2. Protocols for cell cycle–specific FRAP in the absence of drug synchrony. (A) A cell expressing Mcm4-mEm was imaged emerging from mitosis (top). FRAP experiments were conducted on both sister cells ∼5 h later (bottom). (B) A late G1 cell expressing Mcm4-mEm was imaged before a FRAP bleach (Pre-bleach), and at different time points after the bleach to measure recovery. PCNA foci appear between 30 and 60 min, which indicates entry into S phase. (C) A late S phase cell expressing Mcm4-mEm was imaged until all PCNA foci had cleared, followed by a FRAP experiment in the subsequent G2 phase. PCNA foci disappear between 70 and 135 min, which indicated entry into G2 phase. Note that for live cell images, exposure time is minimized to 1/10th of a second, so resolution is compromised. Bars, 10 µm.

Protocols for cell cycle–specific FRAP in the absence of drug synchrony. (A) A cell expressing Mcm4-mEm was imaged emerging from mitosis (top). FRAP experiments were conducted on both sister cells ∼5 h later (bottom). (B) A late G1 cell expressing Mcm4-mEm was imaged before a FRAP bleach (Pre-bleach), and at different time points after the bleach to measure recovery. PCNA foci appear between 30 and 60 min, which indicates entry into S phase. (C) A late S phase cell expressing Mcm4-mEm was imaged until all PCNA foci had cleared, followed by a FRAP experiment in the subsequent G2 phase. PCNA foci disappear between 70 and 135 min, which indicated entry into G2 phase. Note that for live cell images, exposure time is minimized to 1/10th of a second, so resolution is compromised. Bars, 10 µm.

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