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Fig. 1. Immunoblotting analyses and the distribution of epiplakin in cultured cells. (A) Immunoblot of NHEK-L (lane 2) and Cos-7 (lane 3) extracts probed with an antibody to epiplakin. ß-tubulin is shown as a loading control. (B) Equal amounts of the soluble (S) and insoluble (P) fractions were resolved in a 3% SDS-NuPAGE gel and transferred to PVDF membrane, probed with the anti-epiplakin (1:2000) and the HRP-conjugated anti-rabbit antibodies (1:20,000). Nebulin, with a molecular mass of 850 kDa, is marked. (C) Immunofluorescent staining of HeLa (a), A431 (b), HaCaT (c) and NHEK-L (d) cells. Cells were fixed in cold methanol and stained with anti-epiplakin antibody. The thin filament structure is indicated by arrows in d. Bars, 20 µm.





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