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

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posted on 2013-04-03, 06:22 authored 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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