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Fig. 7. Stau2 splice isoforms are found in high-density particles. (A) Rat cortical neurons (6 DIV) were lysed, homogenized and cytoplasmic and nuclear extracts were prepared by low-speed centrifugation. The supernatant (cytosolic fraction, C) and pellet (nuclear fraction, N) were recovered and analyzed by western blotting with monoclonal anti-Stau2 antibodies. The purity of each fraction was tested with antibodies directed against a nuclear (histone H1) and a cytosolic (ribosomal protein P) protein. (B) Western blot analysis of the S100 and P100 fractions following different treatments. Cytoplasmic extracts were either left untreated (-) or were treated for 30 minutes prior to centrifugation with 0.5 M KCl (KCl), 0.5% Nonidet P40 (NP40), 25 mM EDTA (EDTA), 300 U/ml micrococcal nuclease (RNase) and 0.5% Nonidet P40 and 25 mM EDTA (NP40+EDTA). S100/P100 fractions were analyzed with monoclonal anti-Stau2 (Stau2), anti-ribosomal protein L7a (L7a), anti-{alpha}-tubulin (Tub), and anti-calnexin (CNX) antibodies. The same results were obtained in two independent experiments.