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. 2017 Nov;23(11):1660–1671. doi: 10.1261/rna.062000.117

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© 2017 Donovan et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it 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 4. — Northern blot analysis of Y-RNA and tRNA cleavage during dsRNA response. (A) RNA chip analysis for cleavage of total RNA in A549 cells treated with poly(IC). Markers show fragments produced by RNase L. RNA is intact in RNase L^−/− cells. (B) Northern blot analysis for each Y-RNA after 5 h of poly(IC) treatment. RNA fragments are less abundant than the starting Y-RNAs, but detectable, indicating that they survive for some time in the cells. Relative abundance is normalized to RNU6. (C) Northern blot analysis of tRNA cleavage during poly(IC) treatment. Cleavage of tRNA-His and tRNA-Pro are readily detected but only tRNA-His is depleted upon prolonged treatment. (D) Western blot analysis to detect inhibition of new protein synthesis during poly(IC) treatment. Poly(IC) does not change the abundance of preexisting total protein, but blocks translation of new proteins. RNase L is important for early and strong translation arrest. Western blot for eIF2α phosphorylation status reflects PKR activation.