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. 2011 Sep;31(18):3773–3789. doi: 10.1128/MCB.05263-11

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Fig. 2. — Collagen α1(I) and α2(I) mRNAs cofractionate with vimentin filaments. (A) Insoluble vimentin representing filaments (INS; lane 1) and soluble vimentin (SOL; lane 2) were fractionated by detergent lysis and centrifugation. The fractions (FRAC) were probed for the presence of vimentin (VIM), tubulin (TUB), myosin IIB (MYO), actin (ACT), fibronectin (FIB), and LARP6 proteins by Western blotting. (B) Collagen mRNAs segregate with insoluble vimentin. The fractions in panel A were analyzed by RT-PCR for collagen α1(I), collagen α2(I), GAPDH, actin, and fibronectin mRNAs. Shown is pulldown of collagen mRNAs with an antivimentin antibody. (C) Collagen mRNAs in the insoluble fraction coimmunoprecipitate with vimentin. The insoluble fraction (INS) was redissolved, and immunoprecipitation with vimentin antibody (lane 1) or control antibody (lane 2) was performed. (Lanes 3 and 4) The same experiment using the soluble fraction (SOL). The immunoprecipitated material was analyzed by RT-PCR for the presence of collagen α1(I), collagen α2(I), and fibronectin mRNAs.