Skip to main content
. 2016 Jun 6;6:27401. doi: 10.1038/srep27401

Figure 2. Distribution of Pol II populations during mitosis.

Figure 2

(a) The Pol 3.3 monoclonal antibody (mAb) (αRpb1) recognizes a Pol II epitope outside of the CTD and allows the visualization of the distribution of total Pol II in cells (green). Tyr1-P, Ser2-P, Ser5-P, Ser7-P, or Thr4-P-specific mAbs show the abundance and distribution of individual CTD modifications (red) during mitosis. Merged images show localization of signals in the cytoplasm, but also reveal strong enrichment of Thr4-P signals in two distinct foci (red). DAPI: 4′,6-diamidino-2-phenylindole. Representative images of metaphase chromosomes are shown. Data are from three experiments in which at least 100 cells were analyzed and >98% of mitotic cells show these Pol II CTD-P distributions. (b) Images of a Thr4-P-specific mAb (6D7) and DNA (DAPI) in different mitotic human cells (HepG2, H1299). (c) Immunofluorescence images of Thr4-P-specific mAbs of different subclasses (6D7, 1G7, 4H2; red) and a centrosome-specific Ab (γ-tubulin; green) in HeLa cells. Representative images of metaphase chromosomes from three experiments are shown. Signals were merged and quantified using Image J 1.37 V and the plug-in RGB profiler. Line scans were used to measure Thr4-P-specific and γ-tubulin-specific signals. Scale bars, 5 μm.