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. Author manuscript; available in PMC: 2015 Mar 27.
Published in final edited form as: Cell. 2014 Mar 27;157(1):13–25. doi: 10.1016/j.cell.2014.02.009

Figure 5. Emerging imaging technologies.

Figure 5

(A) Single molecule super-resolution imaging of RNA Pol II (Ignacio Izeddin, Ibrahim Cisse, Maxime Dahan and Xavier Darzacq, personal communication). Three-dimensional density map of Pol II localization in fixed nuclei (left) highlights spatial Pol II clustering, while single particle tracking in live cells (right) identifies distinct Pol II dynamic behaviors. Data were collected from an engineered cell line stably expressing the Pol II catalytic subunit (RPB1) labeled with the photo-convertible fluorescent protein Dendra2 (Cisse et al., 2013). (B) Promoter-specific transcription initiation directed by a reconstituted human Pol II system at single molecule resolution using TIRF video-microscopy. Cy5-labeled DNA templates containing a consensus Pol II promoter are immobilized on a surface, and nascent transcripts are detected based on colocalization of fluorescent probes and template signals. The two DNA templates contain (red) or lack (green) the target sequence for the transcript probe to control for specificity (adapted from (Revyakin et al., 2012)).