Supplementary Materials

Supplementary Materials for:

Computational spatiotemporal analysis identifies WAVE2 and cofilin as joint regulators of costimulation-mediated T cell actin dynamics

Kole T. Roybal, Taráz E. Buck, Xiongtao Ruan, Baek Hwan Cho, Danielle J. Clark, Rachel Ambler, Helen M. Tunbridge, Jianwei Zhang, Paul Verkade, Christoph Wülfing,* Robert F. Murphy*

*Corresponding author. E-mail: christoph.wuelfing{at}bristol.ac.uk (C.W.); murphy{at}cmu.edu (R.F.M.)

This PDF file includes:

  • Fig. S1. Example images from the segmentation, alignment, and morphing pipeline.
  • Fig. S2. Illustrations of the SDs of the spatiotemporal models.
  • Fig. S3. Spatiotemporal models for actin distribution using two-point annotation.
  • Fig. S4. Analysis of radial distributions confirms differences between myosin and actin.
  • Fig. S5. Comparison of LAT spatiotemporal dynamics reveals restoration of costimulation by active Rac and cofilin.
  • Fig. S6. Quantification of actin regulatory proteins in primary mouse T cells.
  • Fig. S7. Volume measurements of primary mouse T cells.
  • Fig. S8. Variation in cell shape and morphing across time points and conditions.
  • Table S1. The number of imaging fields and manually marked immunological synapses used to construct the spatiotemporal model of each condition-sensor combination.
  • Table S2. The manually tuned parameters used for both stages of segmentation by the active contour method.
  • Table S3. Average amount of morphing required for each sensor at each time point.
  • Table S4. Statistical testing of differences in sensor enrichment in the immunological synapse region between the full stimulus condition and the costimulation blockade.
  • Legend for movie S1

[Download PDF]

Technical Details

Format: Adobe Acrobat PDF

Size: 4.35 MB

Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.mov format). Four-dimensional maps of three representative sensors.
  • Data file

Citation: K. T. Roybal, T. E. Buck, X. Ruan, B. H. Cho, D. J. Clark, R. Ambler, H. M. Tunbridge, J. Zhang, P. Verkade, C. Wülfing, R. F. Murphy, Computational spatiotemporal analysis identifies WAVE2 and cofilin as joint regulators of costimulation-mediated T cell actin dynamics. Sci. Signal. 9, rs3 (2016).

© 2016 American Association for the Advancement of Science