FIG. 9.

Interaction of the cytoplasmic domains of LPP3 with p120ctn. (A) Sequences of the N- and C-terminal cytoplasmic (cyto) domains of LPP3. (B) Illustrations of the GST-N-cyto (GST-NC) and GST-C-cyto (GST-CC) fusion proteins. (C) Purity of GST (∼29 kDa), GST-CC (∼32 kDa), and GST-NC (∼31 kDa) was verified by SDS-PAGE and Coomassie blue staining. Faster-migrating species represent GST alone and fusion protein degradation products. (D to I) EC extracts were subjected to IP using the indicated antibodies, and the resulting immunoprecipitates were subjected to far-Western analysis by probing the membranes with GST-LPP3-C-cyto and then anti-GST MAb. Individual lanes from the blot in panel D were excised and probed with anti-VE-cadherin MAb (E), anti-p120ctn MAb (F), anti-β-catenin MAb (G), anti-γ-catenin MAb (H), or anti-β₁-integrin pAb (I). (J to M) EC extracts were subjected to GST pulldown with the indicated fusion proteins. Precipitates were analyzed by WB with anti-p120ctn (MAb) (J and K) and anti-GST MAb (L and M). All blots shown are representative of at least three separate experiments.