Lim Lab | Papers

A Polybasic Motif Allows N-WASP to Act as a Sensor of PIP2 Density

Venizelos Papayannopoulos, Carl Co, Kenneth E. Prehoda, Scott Snapper, Jack Taunton, and Wendell A. Lim
Molecular Cell 17, 181-191 (2005)

Supplemental Figure S1. PIP2 binding assays
Supplemental Figure S2. Actin polymerization assays
Supplemental Figure S3. PIP2 activation curve for deltaEVH1-N-WASP
Supplemental Figure S4. Activation of N-WASP by PIP3
Supplemental Figure S5. Co-activation by Cdc42

Timelapse Movies: Endosomal Vesicle Rocketing in Cells Transfected with Wt and 14K N-WASP
Supplemental Movie 1
Supplemental Movie 2

Movies were made from timelapse images taken every 5 s over a time period of 5–10 min. Live cells were visualized by fluorescence microscopy under 600× magnification 24 hr after transfection with the appropriate constructs (N-WASP variant and actin-GFP). Supplemental Movie 1 depicts a cell transfected with 14K N-WASP and actin-GFP. Actin comet tails can be seen rocketing around the cytoplasm. For comparison, Supplemental Movie 2 depicts an example of a cell transfected with 9K N-WASP and actin-GFP. No actin rocketing is observed. Only slight fluctuations of vacuoles are observed. Played with Windows Media Player, these movies run 15 s (14 K) and 8 s (9 K) each. The movies have been accelerated by a factor of approximately 35-fold above real time. See Experimental Procedures for further details.