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. 2009 Oct;15(10):1837–1848. doi: 10.1261/rna.1650109

FIGURE 1.

FIGURE 1.

(A) Lsm1-7–Pat1 complexes isolated from the lsm1-8 and lsm1-6 mutants contain all the component proteins. Complexes purified from the wild-type and mutant cells (indicated above the lanes) were subjected to SDS-PAGE, and the bands were visualized by silver staining. Identity of the proteins present in the various bands (revealed by mass spectrometry analysis) is indicated on the right. (B) Coprecipitation of mRNA with the mutant Lsm1p is almost completely abolished in the case of the lsm1-8 mutant but only partly impaired in the case of the lsm1-6, lsm1-9, and lsm1-14 mutants. Anti-FLAG antibody immunoprecipitations were carried out from the lysates of various strains (indicated above the lanes), after which the RNA extracted from the entire pull-down fraction (middle panel) or a small aliquot of the input lysate (top panel) were subjected to Northern analysis for MFA2pG mRNA. (bottom panel) Western analysis using anti-FLAG antibodies to visualize the FLAG-protein present in the pull-down fractions. The fraction of the lysate mRNA that got coprecipitated in each experiment is indicated as a percentage below the middle panel. Total RNA from wild-type yeast was loaded in the first lane (from left) in the top and middle panels to show the poly(A) tail distribution of the MFA2pG mRNA.