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. 2017 Oct 12;7:13055. doi: 10.1038/s41598-017-12915-1

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Polη affects elongation in vivo and is enriched over the active GAL1 gene (a) Schematic drawing of the G-less based run-on (GLRO) assay. The sizes of the two G-less cassettes are shown. (b) GLRO analysis was performed with the indicated strains transformed with the GLRO-long plasmid. A representative gel picture is shown. See Fig. S8 for the full-length image. (c) Quantification of the results of four independent GLRO experiments. For each sample, the ratio of total counts incorporated into the distal versus the proximal G-less cassettes was normalized to the ratio in the wild-type strain, which was set to 100%. (d) Occupancy of Polη on the UAS, 5′ORF, 3′ORF of the GAL1 gene and on two independent intergenic regions in uninduced (raf) and induced (gal) conditions was measured by chromatin immunoprecipitation (ChIP) using anti-Myc antibody in a strain arrested in G1, and expressing C-terminally Myc-tagged Polη. As control, ChIP was also performed with an untagged strain (no tag). Percentage of input at the indicated regions was normalized to intergenic region 2 on chromosome IV. Experiments were repeated at least 3 times. Mean and standard deviations are indicated, p-values were calculated by 2-tailed t-test, n.s.: no statistical difference.