Dvl clusters Gpr125 and Gpc4/Kny into membrane subdomains and promotes uniform Vangl2/Tri membrane localization in late blastulae (4-5 hpf). (A-B′) Animal pole views of live blastulae co-injected with 267 pg gpr125-Cherry RNA and either 150 pg mEGFP RNA (A-A′) or 150 pg dvl-GFP RNA (B-B2). Arrows in B′ indicate Gpr125-Cherry:Dvl-GFP membrane subdomains. (C-D′) Animal pole views of live blastulae co-injected with 60 pg gpc4/kny-GFP RNA, 50 pg mCherry RNA (C-C′) and 150 pg dvl RNA (D-D′). Arrows in D′ indicate Gpc4/Kny-GFP membrane subdomains. (E) Graphic representation of the relative distribution of Dvl-GFP:Gpr125-Cherry and Gpc4/Kny-GFP:Dvl subdomains along cell membranes. Membrane length was normalized as one. (F,F2) Animal pole views of a whole-mount immunostained wild-type blastula with Vangl2 and β-catenin antibodies. (G-G′) Animal pole views of a 50 pg mCherry and 150 pg dvl-GFP RNA-injected blastula immunostained for GFP, RFP and Vangl2.
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