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. 2003 Jul;23(14):4972–4982. doi: 10.1128/MCB.23.14.4972-4982.2003

FIG. 4.

FIG. 4.

Binding of LRP130 to hnRNP A1-associated RNAs is specific and occurs in vivo. (A) RNP complexes were immunopurified from HeLa cell lysates with antibodies against hnRNP A1 (4B10), hnRNP C1/C2 (4F4), and nucleolin (7G2). Proteins in the isolated complexes were resolved by SDS-PAGE and analyzed by immunoblotting with the indicated antibodies. Five percent of the material used for immunoprecipitation was loaded in lane Total. (B) Cross-linked RNP complexes from HeLa cells exposed to UV light were selected from a Triton X-100-extracted subcellular fraction as described in the text and subjected to immunoprecipitation with antibodies against hnRNP A1 or nucleolin (lanes 5 and 7). Proteins in the immunopurified complexes were released by digestion with RNase, resolved by SDS-PAGE, and analyzed by immunoblotting with an antibody against LRP130. Lanes 2 and 8, lysates from cells that were not exposed to UV light; lanes Total, 1% of the material loaded on the oligo(dT) columns; lanes 4 and 6, mock immunopurifications (IP) in which HeLa cell material was omitted; lanes 3 and 8, total oligo(dT)-eluted material from cross-linked or control cells equal to the amount used for the immunopurifications shown in lanes 5 and 7.