Fig. 2

MAMLD1 confers the nuclear translocation ability of YAP1-MAMLD1 fusion protein. a YAP1 fusion and control constructs at protein level and graphical illustration of the workflow used in this study. Red dashed line indicates fusion site. All constructs are tagged with the human influenza hemagglutinin surface glycoprotein (HA). Color codes are identical to Fig. 1b. The YAP1-MAMLD1 fusion or indicated control constructs were cloned into the pT2K transposable vector and injected with or without the Tol2 transposase into the lateral ventricle of E13.5 wild-type mice followed by transfection using an electroporation based in vivo gene transfer approach. NLS nuclear localization signal. CAG CMV early enhancer/chicken β actin promotor, IRES internal ribosomal entry site, Tol2 Tol2 transposase cis element. b Immunofluorescence micrographs for subcellular localization of indicated proteins in the cells of the ventricular zone 2 days after in utero electroporation. Mock represents the pT2K IRES-EGFP empty plasmid. Double staining was performed for DAPI/HA (left panel) or EGFP/DAPI (mid-panel) or HA/EGFP (right panel) (scale bar, 10 µm). c Quantification of the percentage of the electroporated cells expressing indicated exogenous (HA-tagged) proteins in the nucleus and the cytoplasm. d Immunostaining of LN229 cells co-expressing Myc-tagged Lats2 and indicated genes tagged by HA. The cells were stained with anti-HA and anti-Myc antibodies followed by counterstaining with DAPI. e Western blot of protein lysates of LN229 cells transfected with indicated genes. f Immunoprecipitation of protein lysates of the transfected LN229 cells with the Flag antibody followed by western blotting with indicated antibodies. Actin is used as an internal control