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. 2011 Nov 24;40(6):2527–2539. doi: 10.1093/nar/gkr1040

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© The Author(s) 2011. Published by Oxford University Press.

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

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Figure 4. — Effect of rapamycin and its removal on ribosome distribution. HeLa cells were incubated with 100 nM rapamycin for 4 h and then the medium was replaced with fresh medium without the inhibitor for 2 h before the lysis. (A) Total RNA was extracted and subjected to northern blot analysis for pre-rRNA species. The membrane was hybridized with probes ITS1, ITS2, 18S and 28S as previously described. (B) Polysomal profile after rapamycin removal of HeLa cells. The polysomal and non-polysomal fractions are indicated. (C) Northern blot analysis of cytoplasmic RNA extracts from each fraction of sucrose gradient and analyzed with indicated probes (RpS6, RpS19 and β-actin). (D) Quantification of the signals is reported as a linear plot of percentage of mRNAs in each fraction.