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Figure 2


Fig. 2. LINT-25 and LAP2{alpha} interact in vitro. (A) His-tagged, recombinant LINT-25 and LAP2{alpha} were incubated alone or together and mixed with immobilized LAP2{alpha} antibody. Immunoprecipitates (P) and supernatants (SN) were analyzed by immunoblotting detecting LAP2{alpha} (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 LAP2{alpha} or LAP2beta1-408 and mixed with immobilized LAP2{alpha}- and LAP2beta 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 {alpha}His-tag reagent ({alpha}His) or overlaid with recombinant LAP2{alpha} or buffer. Bound LAP2{alpha} was detected using LAP2{alpha} antibodies ({alpha}LAP2{alpha}). Numbers indicate molecular masses of marker proteins in kDa; mLINT-25, monomeric LINT-25; dLINT-25, dimeric LINT-25.





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