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Fig. 7. Cooperative effects and physical interactions within the Gli superfamily.
(A) (Top left) Human GLI1 is nuclear in transfected COS cells, here detected
with specific polyclonal anti-human GLI antibodies
(Lee et al., 1997). (Top
right) Myc-tagged Gli2 is cytoplasmic. (Bottom left) Co-expression of GLI1 and
Gli2 results in nuclear localization of Gli2 (bottom right). (B) (Top panels)
Expression of Myc-tagged frog Gli1, Myc-tagged-Gli2, Myc-tagged GLI3 and
FLAG-tagged Zic2 in COS cells. (Bottom panels) Co-expression with Zic2 forces
the Gli1, Gli2 and GLI3 cytoplasmic proteins into the nucleus. DAPI staining
highlights nuclei (blue). The color of the font indicates the protein
visualized by immunostaining. (C,D) Quantification of the subcellular
compartmentalization of Gli proteins when transfected alone versus
co-transfection with human GLI1 (C), and when transfected alone versus
co-transfection with Zic2 (D). (C) Only nuclear values are given for
co-transfection of Gli2 and GLI1 and for GLI3 and GLI1. (E-H) (Top panels)
Immunoprecipitation of Flag-tagged proteins with anti-FLAG Ab as labeled,
followed by western blotting with anti-Myc Ab. Arrowheads indicate the
immunoprecipitated species. (Bottom panels) Corresponding western analysis of
total Myc-protein input from transfected COS cells as labeled. The horizontal
alignment of bottom panels corresponds to the molecular weight markers on the
left.
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