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. 2022 May 27;50(11):6453–6473. doi: 10.1093/nar/gkac366

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© The Author(s) 2022. 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 (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — eL39 is stably accommodated into early and medium pre-60S ribosomal particles. (A) eL39 was affinity-purified with GFP-Trap^®_A beads, using total cell extracts of ORY211 (rpl39Δ) cells expressing eL39-yEGFP or untagged eL39 (as a negative control). Total RNA was extracted from whole cell extracts (T) and immunoprecipitates (IP), and analyzed by northern blotting. The indicated probes (in parentheses) were used to detect the different rRNA species (Figure S1A, Table S3). (B) Pre-60S r-subunit assembly intermediates were purified via TAP-tagged AFs Ssf1 and Nog2. The protein composition of the purified particles was analyzed by SDS-PAGE followed by silver staining (left) and western blotting with antibodies against the indicated r-proteins and AFs (right).