Figure 3. Immunofluorescence of membrane and cytoskeletal protein localisation during human erythroblast enucleation.

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. Images shown are slices through cells in early (upper row) and late stages (lower row) of the enucleation process and detected with monoclonal antibodies against alpha spectrin, beta spectrin, ankyrin, band 3, GPC, GPA, RhAG, Rh, CD47 and a rabbit polyclonal antibody against CD44 and a suitable species specific fluorescent secondary as described in materials and methods. N = 5 for each antibody (although generally between 5–20) except for beta spectrin due to problems with high background fluorescence in the nucleus. Scale bar = 5 µm.