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. 2009 Dec 1;23(23):2742–2752. doi: 10.1101/gad.1820709

Figure 1.

Figure 1.

Purification of Vas–mRNA complexes. (A) Aliquots (1/100) of eluates from oligo(dT) chromatography were resolved on a 0.75% TAE agarose gel, blotted, and treated with anti-Vas antibody. (Lanes 1–3) First, second, and third eluates from uncross-linked controls. (Lanes 4–6) First, second, and third eluates from cross-linked samples. (Lane 7) Cross-linked lysate before oligo(dT) chromatography, a positive control. (B) Purification of Vas–mRNA complexes by tandem Vas immunoprecipitation, analyzed by immunoblot as in A. (Lane 1) Cross-linked sample before immunoprecipitation, a positive control. (Lanes 2,3) Aliquots of material recovered from the first immunoprecipitation, using rabbit IgG (control, lane 2) and α-Vas (lane 3). (Lanes 4,5) Aliquots of material recovered from the second immunoprecipitation using rabbit IgG (control, lane 4) and α-Vas (lane 5). The arrows indicate the purified Vas–mRNA complexes. (C) Synthesis of double-strand cDNA. Lane 1 is a positive control. Lanes 2–4 are the first-strand cDNA synthesis, and represent buffer control (lane 2), uncross-linked control (lane 3), and cross-linked sample (lane 4). Lanes 5–7 are the second-strand cDNA synthesis, and represent buffer control (lane 5), uncross-linked control (lane 6), and cross-linked sample (lane 7).