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. 2001 Jul 9;154(1):71–84. doi: 10.1083/jcb.200101089

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Copyright © 2001, The Rockefeller University Press

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Figure 2. — Turnover of NPCs and the lamina measured by FRAP. (A) FRAP of NRK_POM121-YFP3 cells. The boxed area was photobleached to background levels. Recovery was monitored immediately after the bleach and every 30 min by taking a stack of five confocal images after autofocussing. Representative maximum intensity projections are shown. Arrowheads mark the boundary between bleached and nonbleached regions. Time, hh:mm:ss. See Video 1 for the entire sequence. (B) FRAP experiment similar to A of NRK cells transiently expressing EGFP–lamin B1. Single confocal sections 0.5 μm above the coverslip surface were acquired every 15 min after autofocussing. Times, scale, and arrowheads as in A. See Video 2 for the entire sequence. (C) Plot shows fluorescence recovery of POM121 (green) and lamin B1 (black) in the bleached regions. Bleached and nonbleached half of the nuclei were tracked manually to measure mean intensities. Values were background subtracted and then normalized to total loss of fluorescence. Bars, 5 μm. Online supplemental material (Videos 1 and 2) is available at http://www.jcb.org/cgi/content/full/200101089/DC1.

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