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. Author manuscript; available in PMC: 2010 Nov 1.
Published in final edited form as: Biochim Biophys Acta. 2009 Aug 27;1793(11):1787–1794. doi: 10.1016/j.bbamcr.2009.08.009

Fig 5. Co-precipitation of LRBP and LH receptor mRNA in the oligo(dT) eluted mRNP fraction.

Fig 5

Fig 5

Fig 5

mRNA-binding proteins in the polysomes from 0 h control (CTL) and downregulated (hCG) pseudopregnant rat ovaries collected 2, 4, 6 and 12 h after hCG treatment were separated using oligo(dT) cellulose chromatography as described in detail in Materials and Methods. Proteins from the oligo(dT) cellulose bound mRNP complexes were eluted using SDS sample buffer and subjected to Western blot analysis using LRBP antibody (A). The blot shown is a representative of three independent experiments. Oligo(dT) cellulose bound mRNPs were eluted using 10 mM Tris/HCl, pH 7.5 and subjected to immunoprecipitation using LRBP antibody. The immune complex was reverse transcribed and the cDNAs were used for real time PCR using pre-designed primers and probes for rat LH receptor mRNA (B). Total RNA was also extracted from the mRNP eluate directly and was reverse transcribed and subjected to real time PCR analysis using pre-designed primers and probes for rat β–actin, GAPDH and RPS6 (C). The graphs represents changes in mRNA levels normalized to 18S rRNA, and are shown as fold changes vs. control. Error bars represent mean ±SE. *p<0.05, n=3.