Fig. 5.

The Wnt agonist LiCl stimulates formation of endogenous active Rac1–β-catenin complexes in the cytoplasm and nucleus. In situ PLA in HEK293T and NIH 3T3 cells after Wnt stimulation with LiCl. (A) Cells were treated with 40 mM LiCl for 6 h and antibodies against active Rac1 and β-catenin were used to detect active Rac1–β-catenin complexes. The primary antibodies Rac1-GTP and β-catenin were used alone as controls (top two rows). Cells were counterstained with anti-β-catenin (green) and Hoechst (blue). (B) Positive PLA signals (red dots) were counted from z-stack confocal images and the number of dots in the nuclei of cells is presented in the dot plots for each cell line. Fifty nuclei per experiment were counted over two independent experiments and a representative plot is displayed. Unt, untreated.