FIG. 7.

An LS mutant enhances ANF expression and promotes cardiac cell hypertrophy through a PI3K-dependent pathway. (A) Myocardial cushions from chicken embryos were infected with adenoviruses expressing GFP and a SHP2-WT, or -T468M mutant. After stimulation with EGF, the sizes of cushion cells were measured by FACS. (Left) FACS profile from a representative experiment. (Right) Medians of FACS profiles from 3 different experiments. The error bars indicate SEM. (Inset) Confocal images of GFP and rhodamin-phalloidin staining showing tissue integrity at the end of the experiment (left) and GFP staining of a whole cushion infected with SHP2-WT adenoviruses (right). (B) Neonatal rat cardiomyocytes were cotransfected with the indicated SHP2 construct (WT, N308D, or T468M) and a luciferase (Luc) construct under the control of the ANF promoter. Two days after transfection, the cells were assayed for Luc activity. When indicated, the cells were treated, before the Luc assay, overnight with LY294002 (control, cotransfection of ANF-Luc and empty vector). The graphs are presented as the mean and SEM of 3 independent experiments performed in duplicate. In panel A, significant differences versus stimulated WT cells are indicated. In panel B, significant differences versus the untreated control are indicated. (*, P < 0.05; **, P < 0.01; n.s., not significant.)