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. 2013 Dec;19(12):1669–1683. doi: 10.1261/rna.038356.113

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© 2013 Al-Jubran 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 3.0 Unported), as described at http://creativecommons.org/licenses/by-nc/3.0/.

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FIGURE 8. — Puromycin incorporation at transcription sites. (A) Direct immunostaining of polytene chromosomes with Alexa488-conjugated anti-puromycin antibody. Top panels show salivary glands pre-incubated with puromycin and emetine, and the bottom panels show glands incubated with puromycin, emetine, and actinomycin D. Details are in Materials and Methods. Lower insets show higher magnification DAPI images of the regions indicated, and the upper insets are merged fluorescence and DAPI images. Arrows indicate interbands/puffs with strong anti-puromycin fluorescence; the pointer indicates a fluorescent band associated with a condensed chromosomal region. (B) Immunostaining of chromosomes as in A, of glands treated with RNase A (100 µg/mL for 2 min at room temperature to the first squashing step, before fixation).