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. 2010 Nov 24;39(7):2671–2677. doi: 10.1093/nar/gkq1190

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© The Author(s) 2010. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Cyclin E is present in nuclear matrix-associated foci. (A) Immunofluorescent detection of cyclin E (green) and DNA (blue) in cycling NIH3T3 cells after extraction with or without DNase. Bar is 5 μm. Histogram shows average fluorescence intensity derived from multiple images with SEM. (B) Western blot showing cyclin E and other proteins as indicated, in fractions generated by serial extraction of NIH3T3 cells. T = total protein. Cells were lysed in detergent containing buffer to generate pellet (P) and supernatant (SN). Pellet was washed with 0.5 M NaCl to generate ‘w’ and extracted with or without DNase. All fractions are derived from cell equivalents. (C and D) As in B, showing cyclin E in early passage murine cardiac fibroblasts and human lung cell line HFL1, respectively. (E) The proportion of NIH3T3 cells engaged in DNA synthesis between 14 and 19 h after release from quiescence, measured by incorporation of BrdU. Inset shows BrdU (red) in nuclei (blue) at 19 h. (F) Western blots showing total cyclin E, CDC6 and histone H3 14–19 h after release from quiescence (upper panel). After extraction with DNase cyclin E remained in the pellet (middle panel), despite complete release of histone H3 (lower panel), at all time points.