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. 2015 Feb;21(2):202–212. doi: 10.1261/rna.048199.114

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© 2015 Alings et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article, published in RNA, is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 5. — Inhibition of tRNA synthesis stalls 2-thiolation. (A) Growth curve measurements of S288C cells in logarithmic growth at 37°C supplemented with RNA polymerase inhibitor ML-60218 (top panel) or thiolutin (middle panel) at the 0 h time point. rpo31-698 was grown at 30°C and shifted to 37°C at the 0 h time point (bottom panel). (B) Representative autoradiogram of tRNA extracted at indicated time points from ML-60218 treated cells following APM-affinity gel electrophoresis and Northern blot with tE^UUC probe. Arrowheads highlight the 0 h time point at which the inhibitor was added. Thiolation levels were quantified by image densitometry (n = 3). (C) As in B for thiolutin. (D) As in B for rpo31-698, where inhibition of tRNA synthesis is induced by the temperature shift to 37°C.