A Targeted siRNA Screen Identifies Regulators of Cdc42 Activity at the Natural Killer Cell Immunological Synapse

Leo M. Carlin,* Rachel Evans, Hanna Milewicz, Luis Fernandes, Daniel R. Matthews, Michela Perani, James Levitt, Melanie D. Keppler, James Monypenny, Ton Coolen, Paul R. Barber, Borivoj Vojnovic, Klaus Suhling, Franca Fraternali, Simon Ameer-Beg, Peter J. Parker, N. Shaun B. Thomas, Tony Ng*

*To whom correspondence should be addressed. E-mail: leo.carlin{at}kcl.ac.uk (L.M.C.); tony.ng{at}kcl.ac.uk (T.N.)

This PDF file includes:

• Fig. S1. Diagram of the Raichu-Cdc42 biosensor and time-lapse live-cell FLIM analysis of a representative NK cell containing Cdc42-GFP.
• Fig. S2. Cdc42 protein interaction network and iPFAM scoring method.
• Fig. S3. Quantification of siRNA-mediated knockdown and Raichu-FLIM for fixed cell inhibitor experiments.
• Table S1. Table of PubMed IDs for interaction evidence used to prepare the interaction subnetwork.
  

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Other Supplementary Material for this manuscript includes the following:

• Movie S1 (.mov format). Time-lapse live-cell Raichu-FLIM movie showing a YTS NK cell interacting with two 721.221 target cells, including an animated plot of the mean FRET efficiency of Raichu-Cdc42.

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© 2011 American Association for the Advancement of Science