Fig. 2.
PAK6 targets to cell–cell adhesions independent of catalytic activity through its N-terminus. (A) Schematic representing generated GFP-tagged PAK6 constructs. (B) 5.0×105 cells expressing the GFP-tagged PAK6 constructs were lysed directly in sample buffer and immunoblotted for GFP to indicate exogenous constructs are the expected molecular weights. (C,D) Stable shPAK6 DU145 cell lines expressing the indicated constructs were plated on glass coverslips, fixed after 24 h and stained for F-actin (phalloidin) or β-catenin. (E) Mean percentage (±s.e.m.) of cells in which GFP-tagged constructs targeted to cell–cell adhesions. n=3 with ≥40 cells per experiment. (F) Mean Manders' coefficient (±s.d.) of cells with GFP-tagged constructs targeted to β-catenin positive cell–cell adhesions (from at least 10 fields, >60 cells per condition). Scale bars: 10 µm.