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Facultative heterochromatin is cell-type specific, well documented by electron microscopy and found lining the lamina at the inside of the nucleus.
Analysis of the localization of TaSUT2 at the subcellular level through transient expression of a TaSUT2-YFP fusion protein in onion epidermal cells revealed strong YFP-fluorescing signal on the inside of the nucleus around a structure that appears to be the vacuole, indicating that TaSUT2 is localized to the tonoplast rather than to the plasma membrane.
As a postdoc in Ron Vale's lab at UCSF, Ankur discovered that RNA sequences that contain repeated cytosines, or Cs, and guanines, or Gs, can actually stick together and form membraneless blobs or "RNA gels" inside of the nucleus.
Elongation of MTs inside of the nucleus induces tether formation only at the end(s) lacking SPBs (compare Fig. 1, panels c g to k, l, n): during interphase in which the unduplicated SPB is in the cytoplasm (Fig. 1, a, h), ned1 overexpression [6] induces formation of one or two tethers (Fig. 1, k,l).
Oka et al. have now tracked how these proteins are distributed inside of the nucleus, and examined which part of the DNA they bind to, in more detail.
Using ImageJ (ImageJ 1.43U), signal intensity of each CENP-A or CENP-B spot inside of the nucleus (as defined by the DAPI staining) was extracted.
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Immunofluorescence microscopy revealed that the majority of fusion proteins resided in the cytoplasm of cells, while a minor fraction of Tpk2HA was also detectable at the inside rim of the nucleus.
Resulting z-stack images were de-convolved using a constrained iterative algorithm from SlideBook 5.0 software and nuclear volumes were measured by masking the inside of each nucleus, which were delineated by the GFP signal.
The experiments identified another protein called Slx9 that shuttles rapidly between the inside and the outside of the nucleus.
We also assumed that the nucleus is homogeneous inside and the susceptibility of the nucleus is uniform.
DNA never leaves the nucleus of complex cells, but RNA can be found both inside and outside of the nucleus.
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