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Fig. 9. Usherin binding to type IV collagen may facilitate stable integration of usherin in the basement membrane. (A) Basal lamina was extracted from normal (C) and Alport (A) testes. Solubilized proteins were subjected to immunoprecipitation with anti-usherin antibodies and a western blot probed with anti-type IV collagen {alpha}3, {alpha}4 and {alpha}5 antibodies (lanes 1 and 2); immunoprecipitated with anti-type IV collagen {alpha}1 and {alpha}2 antibodies and a western blot probed with anti-usherin antibodies (lanes 3 and 4); or the crude extract analyzed directly (by western blot and probed with anti-usherin antibodies (lanes 5 and 6). Molecular mass markers are in kilodaltons. (B) The experiment in A was run in triplicate using three different sets of animals, and the chemiluminescence signal was quantified and analyzed statistically as described in Materials and Methods. The quantitative data shown in B correspond to the lanes in A.