Supplementary Materials

Time-resolved structured illumination microscopy reveals key principles of Xist RNA spreading

Lisa Rodermund, Heather Coker, Roel Oldenkamp, Guifeng Wei, Joseph Bowness, Bramman Rajkumar, Tatyana Nesterova, David Miguel Susano Pinto, Lothar Schermelleh, Neil Brockdorff

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

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  • Supplementary Text
  • Figs. S1 and S12
  • Tables S1 and S2
  • Captions for Movies S1 and S2
  • References
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Images, Video, and Other Media

Movie S1
3D volume rendering of Xist territory acquired with 3D-SIM. Mouse ES cell labelled with Xist RNA-FISH (magenta) and BglG-Halo-Xist detected with diAcFAM HaloTag ligand (green), corresponding to Fig. 1B.
Movie S2
Live cell imaging of BglG-Halo-Xist. Time series of BglG-Halo-Xist labelled with JF646 HaloTag ligand, acquired with 10 s intervals, corresponding to fig S2E. For each time point 3DSIM z-stacks of 375 raw images (25 z-sections, 5 phases, 3 angles) were recorded. Left panel shows (pseudo)widefield subjected to iterative deconvolution, right panel 3D-SIM with MC filtering applied to reduce reconstructed noise artifacts as described in detail in fig. S3B and Materials and Methods. Lateral (top) and orthogonal (bottom) maximum intensity projections are displayed in false color (Fire LUT, Fiji). Scale bar: 2 μm.