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. 2019 Sep 12;47(20):10852–10864. doi: 10.1093/nar/gkz767

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© The Author(s) 2019. 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/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Has1 is present in multiple copies in 90S pre-ribosomes. (A) Western blot analysis of Has1-FTP purification from the yeast strain expressing both 3HA-Has1 and Has1-FTP using anti-HA, anti-FLAG and anti-Arc1 antibodies respectively as indicated. The change in the molecular size of Has1-FLAG is due to the TEV cleavage during tandem affinity purification. (B) Chart showing the average fold enrichment of 3HA-Has1 and Has1-FLAG based on the quantification of data from two independent experiments. (C) Sucrose density gradient analysis of the FLAG eluate from affinity purification using the strain expressing both Has1-GST and Has1-FTP. Proteins isolated from each fraction were resolved by SDS PAGE and detected by Coomassie staining. (D) Western analysis of the fractions using the anti-Has1 and anti-GST antibodies. (E) Northern analysis of RNA in each fraction. The fractions corresponding to different pre-ribosomes based on the pre-rRNA content are indicated below the image.