Supplementary Materials

Real-time imaging of DNA loop extrusion by condensin

Mahipal Ganji, Indra A. Shaltiel, Shveta Bisht, Eugene Kim, Ana Kalichava, Christian H. Haering, Cees Dekker

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Figs. S1 to S17
  • Captions for movies S1 to S10
  • References

Images, Video, and Other Media

Movie S1
Movie showing homogenously distributed SxO fluorescence intensity along the length of a double-tethered λ-DNA molecule in the absence of condensin.
Movie S2
Movie showing DNA compaction by condensin. Compaction can be identified as a bright fluorescence spot within the SxO-stained DNA.
Movie S3
Movie showing DNA that has been compacted by condensin. Compaction can be identified as a bright fluorescence spot within the SxO-stained DNA. During this movie, a perpendicular flow is applied to reveal that the bright spot constitutes an extended DNA loop.
Movie S4
Movies showing DNA loop extrusion on SxO-stained DNA under a constant flow. In both movies, DNA initially displays an inverted U shape due to the applied flow. As the movies proceed, a bright fluorescence spot appears that grows into an extended loop, which finally stalls. At its maximum size, the DNA molecule appears as an inverted Y shape.
Movie S5
Movies showing DNA loop extrusion on SxO-stained DNA under a constant flow. In both movies, DNA initially displays an inverted U shape due to the applied flow. As the movies proceed, a bright fluorescence spot appears that grows into an extended loop, which finally stalls. At its maximum size, the DNA molecule appears as an inverted Y shape.
Movie S6
Movie that shows single-step disruption of the DNA loops at the end of movie S5, resulting in the return of a homogenous DNA intensity that was identical to the bare DNA at the start of the experiment..
Movie S7
Movie showing condensin-mediated DNA compaction under constant flow perpendicular to DNA. Near the end of the movie, the DNA loop splits, most likely due to a photo-induced double strand break. This rare event shows the two DNA strands that form the extruded loop.
Movie S8
Movie of DNA loop extrusion on double-tethered DNA in the absence of flow. The DNA initially exhibits a homogenous fluorescence intensity along its length. As time progresses, a bright fluorescence spot appears that migrates towards one of the two ends of the attached DNA.
Movie S9
Movie with overlaid channels of fluorescence from SxO-stained DNA (green) and ATTO647N-labeled condensin (red), showing condensin localization to the stem of the extruded DNA loop.
Movie S10
Movie with overlaid channels of fluorescence from SxO-stained DNA (green) and ATTO647N-labeled condensin (red), showing the correlative spatial fluctuations of the ATTO647N condensin signal and the SxO DNA loop.