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. 2017 Aug 25;45(18):10824–10836. doi: 10.1093/nar/gkx734

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© The Author(s) 2017. 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-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 1. — Sucrose gradient fractionation of total extracts from a TetO7-RIO1, NOB1-FPZ strain grown in YPD or in YPD plus doxycycline for 16 h to repress RIO1 expression. (A) Recording of A254 nm profiles. Top: Rio1p-expressing cells (- Dox); bottom: Rio1p-depleted cells (+ Dox). The peaks of free proteins, free 40S and 60S subunits, 80S ribosomes and polysomes are indicated as well as fraction numbers. (B) Sedimentation profile of pre-40S particle AFs analyzed by western blot. Proteins were purified from the indicated fractions (top) of gradients loaded with extracts from Rio1p-expressing cells (–Dox) or Rio1p-depleted cells (+Dox). The antibodies used for the detection of specific factors are indicated. The anti-FLAG antibody allows the detection of tagged Nob1p. (C) Sedimentation profile of (pre)-rRNAs analyzed by northern. Same as in (B) except that RNAs were extracted from gradient fractions, separated by agarose gel electrophoresis in denaturing conditions, transferred to a nylon membrane and detected using various specific oligonucleotide probes.