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. 2021 Oct 22;12:6153. doi: 10.1038/s41467-021-26207-w

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 8 — a Immunoprecipitation of Noc1-TAP-containing preribosomal particles from wild-type, snr190-[mut.C] and dbp7Δ mutant strains, and analysis of their RNA and protein contents by northern and western blotting, respectively. Particles were immunoprecipitated from total cell extracts prepared from wild-type, snr190-[mut.C], and dbp7Δ strains expressing a TAP-tagged version of Noc1. Pre-rRNA components of 90S and early pre-60S r-particles (indicated on the left) present in the total extracts (Inputs) or in the immuno-precipitates (IPs) were analyzed by northern blotting (upper panel) using a radiolabeled probe (Supplementary Table 4). Protein levels of components of the Npa1 complex (indicated in grey) or other proteins present in early preribosomal particles (labeled in black) were analyzed by western blotting on the Input or IP samples (lower panel) using specific antibodies. *: nonspecific signal detected with the anti-Nop8 antibodies. **: Prp43 signal resulting from incubation of the membrane with anti-Prp43 antibodies (see above panel) prior to incubation with anti-Dbp6 antibodies. b Quantification of the western blot signals obtained in a. Western blot signals were quantified from ChemiDoc images (Biorad) using the Image Lab software (Biorad). The histogram represents the co-immunoprecipitation efficiencies (IPs over Inputs) of the indicated proteins with Noc1-TAP in the wild-type (WT, gray), snr190-[mut.C] (black), and dbp7Δ (red) strains, normalized with respect to the ratios in the wild-type strain. Note: the Dbp6 signal obtained in a could not be quantified accurately due to the proximity of the Prp43 signal.