Nuclear envelope tissue culture cells

Fig. 3 Isolation of nuclei from suspension tissue culture cells. (A) Low-power SEM image of nucleus isolated from K.S62 tissue culture cell, after isolation and Percoll density-gradient purification. This particular nucleus shows surface indentations due to its position in the cell cycle (approaching prophase), but the overall surface morphology of the nuclear envelope is largely free of cytoplasmic contamination, and numerous pore complexes (arrowed) arc apparent. Scale bar, 1.0 /u,m. (B) Detail of A, showing cytoplasmic surface of the nuclear envelope with clearly visible nuclear pore complexes, and a typical density of ribosomes (remember the outer nuclear envelope is the innermost element of the rough endoplasmic reticulum). Scale bar, 125 nm.

Nixon I don t think cultured cells are like atherosclerotic plaque cells, from what we have seen. In the data you showed on the expression of different channels, the expression and localization of InsP3 receptors is very different. For example, the InsP3 receptor expression around the nuclear envelope is not seen in tissue. 

In this procedure cell monolayers are detached from culture vessels by trypsin-ization or scratching using a rubber policeman and pelleted by centrifugation. Isolated nuclei and nuclear envelopes are pelleted by centrifugation. From whole organs such as rat liver, tissue cubes of about 1 mm are dissected. The pellets or tissue cubes are then treated in the following manner ... 

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