Figure 1

Subcellular localization of PELO at the cytoskeleton. A, Competition assay to determine the specificity of PELO antibodies. Blots with testis and lung protein extracts were incubated with anti-PELO antibodies in presence (+) or absence (-) of the GST-PELO fusion protein. B, Immunoblot with cellular extracts from different tissues was probed with anti-PELO antibodies. C, PELO is distributed in cytoskeletal and membrane fractions. Immunoblots with cellular extracts from different tissues were probed with anti-PELO antibodies. Blots containing samples of cytoskeletal (CS), cytoplasmic (C), nuclear (N) and membrane (M) fractions from HeLa cells were subsequently probed with polyclonal anti-PELO, monoclonal anti-α-actinin, anti-phospho H3 histone and anti-α-tubulin. D, Co-localization of the PELO antigen and actin stress fibers. Fibroblasts were immunolabeled with PELO antibodies (i, iv) or FITC-phalloidin (ii, v). PELO immunostaining was localized at the cytoskeleton (white arrows), at some regions of plasma membrane (yellow arrowhead), and near nuclear membranes (white arrowhead). The merged images (iii, vi) show that PELO largely co-localized with actin-stress fibers. Scale bars = 20 μm.