LINT-25 and LAP2α interact in vitro.

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LINT-25 and LAP2α interact in vitro. LINT-25 and LAP2α interact in vitro. (A) His-tagged, recombinant LINT-25 and LAP2α were incubated alone or together and mixed with immobilized LAP2α antibody. Immunoprecipitates (P) and supernatants (SN) were analyzed by immunoblotting detecting LAP2α (upper panel) and the His-Tag (lower panel). Input represents 10% of protein used for IPs. (B) In vitro translated, [35S]methionine-labeled LINT-25 was incubated alone or together with in vitro translated and labeled LAP2α or LAP2β1-408 and mixed with immobilized LAP2α- and LAP2β antibodies (IP) or hybridoma medium (DMEM) as a control. Immunoprecipitates (P) and supernatants (SN) were analyzed by SDS-PAGE and autoradiography. Input represents 2% of protein used for IP. Note, that the signals of the different protein bands do not reflect the relative protein levels due to different numbers of methionines in the proteins' primary sequences. (C) Recombinant LINT-25 was transblotted to nitrocellulose, stained with Ponceau S (Ponc), or detected with αHis-tag reagent (αHis) or overlaid with recombinant LAP2α or buffer. Bound LAP2α was detected using LAP2α antibodies (αLAP2α). Numbers indicate molecular masses of marker proteins in kDa; mLINT-25, monomeric LINT-25; dLINT-25, dimeric LINT-25. Nana Naetar et al. J Cell Sci 2007;120:737-747 © The Company of Biologists Limited 2007