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. 2014 May 13;42(11):7330–7345. doi: 10.1093/nar/gku357

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© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — Immunoprecipitation experiments to assess the interactions of Gno1p-HA with specific components of different pre-ribosomal particles. (A) Interactions of Gno1p-HA with various pre-rRNAs. Gno1p-HA was precipitated from yeast extracts using anti-HA antibodies. Total RNAs were extracted from the pellets obtained following immunoprecipitation (lanes IP) or from 1/15th of the total input extracts used for precipitation (lanes Tot.), separated on a 1.2% agarose gel in denaturing conditions, transferred to a nylon membrane and detected using various specific oligonucleotide probes. The efficiency of precipitation (expressed as percentage of input) of a given pre-rRNA was determined using phosphorimager quantification and is indicated to the right of each panel. (B) Interactions of Gno1p-HA with protein components of various pre-ribosomal particles. Immunoprecipitation experiments have been carried out using IgG sepharose and extracts from cells expressing Gno1p-HA and, when indicated, a TAP- or ZZ-tagged component of a subset of pre-ribosomal particles. Proteins have been extracted from the pellets obtained following immunoprecipitation (lanes IP) or from 1/320th of the total input extracts used for precipitation (lanes Tot.) and analyzed by western as described in the legend of Figure 1A.