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. 2008 Apr 4;82(4):959–970. doi: 10.1016/j.ajhg.2008.02.017

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© 2008 The American Society of Human Genetics. Published by Elsevier Ltd. All right reserved..

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Interaction of Nephrocystin-3 and Inversin

(A–C) V5-tagged inversin was coexpressed with N-terminally FLAG-tagged full-length nephrocystin-3 (Flag.NPHP3 FL) or FLAG-tagged CD2AP protein (Flag.CD2AP) as a negative control. Expression of V5-tagged inversin in cellular lysates was confirmed by immunoblotting with V5 antibodies (A). After immunoprecipitation with FLAG antibody, coprecipitating V5-tagged inversin was detected with V5-specific antiserum (B). As control, FLAG-specific antibodies detected precipitated Flag.Inversin and Flag.CD2AP (C).

(D–F) In the reverse experiment, V5-tagged full-length nephrocystin-3 (V.5NPHP3 FL) was coexpressed with Flag-tagged inversin or FLAG-tagged CD2AP. The additional approximately 60 kDa band visible in all lanes (B, C, E, and F) represents the heavy chain. The 100 kDa bands that are visible in (E) are nonspecific bands that also appeared upon long exposure time in (B).