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Immunofluorescence microscopy confirms that band 3 and ER are lost during nuclear extrusion.

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posted on 03.04.2013, 06:22 by Amanda J. Bell, Timothy J. Satchwell, Kate J. Heesom, Bethan R. Hawley, Sabine Kupzig, Matthew Hazell, Rosey Mushens, Andrew Herman, Ashley M. Toye

Human orthochromatic erythroblasts undergoing enucleation after 144 h of differentiation were removed from culture, fixed in 0.5% acrolein and permeabilised using 0.05% Triton X-100. A) Confocal section of enucleating cells labelled with PDI and band 3 antibodies. B) Confocal section showing extruded nuclei (marked with arrows) labelled with band 3 (BRIC170). C) Confocal section of an erythroblast in the late stage of enucleation co-labelled with ankyrin and band 3 antibodies. The non-association of band 3 with ankyrin was observed in every cell identified at the late stage of enucleation (n = 10). Scale bar represents 5 µm.

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