Figure 6: Amotl1 binds Fat4 and promotes cell proliferation.
From: Amotl1 mediates sequestration of the Hippo effector Yap1 downstream of Fat4 to restrict heart growth
(a) Immunodetection of Yap1 localization in primary cultures of cardiomyocytes infected with the indicated adenoviruses. The inset shows co-localization with Ad-HA-(nls)3-Amotl1 in nuclear spots. (b) Increased percentage of proliferating Ki67-positive cardiomyocytes, as counted by flow cytometry from primary cell cultures infected with nuclear Amotl1 (Ad-HA-(nls)3-Amotl1, n=8), compared with controls (Ad-GFP, n=8). Treatment with Yap1 siRNA (n=8) is inhibitory. ***P<0.001 (analysis of variance). (c) Immunoprecipitation (IP) of Amotl1-HA from HEK293 cells transfected or not with Amotl1-HA or Fat4-ΔECD-Flag. IB, immunoblot with the indicated antibodies. (d) Co-localization of Amotl1-HA, Fat4-ΔECD-Flag and endogenous Yap1 in primary cultures of transfected cardiac cells. The inset shows an enlargement of the boxed region. Ctrl, control; ns, no significant difference. Scale bars: 10 μm.
