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For measuring chromatin density, we fused histone H2A with the monomeric Red Fluorescent Protein 1 (H2A-mRFP) and transfected it into various human epithelial cell-lines.
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As with the hESC experiments, for each HELA nucleus selected (N = 10) we measured chromatin movement in three regions and produced for each a time series of the Hoechst 33342 intensity (chromatin density) along the line scan.
As with the hESC and HELA experiments, for each NIH3T3 nucleus selected (N = 10), we measured chromatin movement in three regions and produced for each a time series of the Hoechst 33342 intensity (chromatin density) along the line scan.
Nissim-Rafinia, M. & Meshorer, E. Photobleaching assays (FRAP & FLIP) to measure chromatin protein dynamicsin living embryonic stem cells.
10.1186/s13072-015-0038-0 Overview of all measured parameters for a comparative topology in relation to chromatin density maps.
The access of immunoglobulins required for immunodetection with an estimated size of about 20 nm [ 31] may in addition be hampered when IC channels fall below a critical throat size of 15 20 nm [ 32] (an overview on all measured parameters for a comparative topology in relation to chromatin density maps is provided in Additional file 6).
Labeling whole chromatin with either fluorescent histones or with DNA dyes and measuring the intranuclear steady-state distribution of injected dextrans relative to chromatin density provided evidence for a high degree of penetration of the probes into chromatin (7– 9).
Thus, D delivers an excellent qualitative and quantitative measure of the experimentally obtained spatial correlation of chromatin density.
Here however, we present a new tracking-based method which can detect chromatin density movement and quantify the mechanical dynamics of chromatin in live cells.
One promising approach to quantify chromatin organization and function is through the spatial correlation of chromatin density distribution and, in particular, its fractal dimension [ 17- 20].
Poorey, K. et al. Measuring chromatin interaction dynamics on the second time scale at single-copy genes.
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