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. 2011 Jun 3;30(13):2557–2568. doi: 10.1038/emboj.2011.178

Figure 8.

Figure 8

The carboxy-terminal regulation motif of PTEN controls β-arr1 binding and effects on migration. (A) U373 cells at the leading edge of wounds were microinjected with plasmids encoding β-arr1–YFP; PTEN-G129E; PTEN-C124S; C124S-T383A; C124S-A4; β-arr1–YFP and C124S-T383A or β-arr1–YFP and C124S-A4, and left to migrate overnight. Cells expressing relevant constructs were detected following staining with a polyclonal anti-Myc or anti-PTEN antibody. All cells were observed by staining with rhodamine- or Alexa488-conjugated phalloidin and DAPI. The red dotted line indicates the wound edge. The number of cells that were still at the wound edge following migration overnight was quantified. (B) Graph showing quantification of U373 cells found at the edge of the wound after migration overnight. Between 150 and 200 cells were counted for each condition. Data shown represent mean±s.e.m. of 3–6 independent experiments. (C) Western blot of wild-type PTEN, C124S-T383A or C124S-A4 contained in β-arr1 immunoprecipitates. Data shown represent the mean±s.e.m. calculated from four independent experiments. (D) Cells expressing indicated constructs were detected as in Figure 7A. The red dotted line indicates the wound edge. The number of cells that were still at the wound edge following migration overnight was quantified.