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. Author manuscript; available in PMC: 2014 Sep 24.
Published in final edited form as: Nat Commun. 2014 Mar 24;5:3516. doi: 10.1038/ncomms4516

Figure 1. EphrinB2 is associated and co-localized with flotillin-1.

Figure 1

(a) Flotillin-1 is associated with ephrinB2. Co-immunoprecipitation (Co-IP) and Western analysis of Xenopus oocytes exogenously expressing ephrinB2-Flag with flotillin-1a-HA or -1b-HA. (b) Endogenous ephrinB2 interacts with endogenous flotillin-1. Endogenous ephrinB2 was immunoprecipitated with anti-goat ephrinB2 antibody in HT-29 colon carcinoma cells, and flotillin-1 was detected using anti-mouse flotillin-1 antibody in ephrinB2 immune complexes by Western blotting. (c) Flotillin-1 immunostaining of embryo sections. Immunostaining of endogenous flotillin-1 in stage 15 embryos, and a magnified view of the neural plate region. Embryos were also injected with F1aMO and F1bMO as indicated, and immunostained for flotillin-1. Scale bar represents 200 µm. (d) Sub-cellular localization of flotillin1b-HA and ephrinB2-Flag. Flotillin1b-HA and ephrinB2-Flag RNA was injected into Xenopus embryos and stained with anti-HA or anti-Flag antibodies. Scale bar represents 20 µm. (e) Co-localization of ephrinB2-mCherry and flotillin-1-GFP at the plasma membrane in embryos. Flotillin-1 GFP (green) and ephrinB2-mCherry (red) RNAs were injected into embryos and localization of proteins was observed by confocal microscopy. The merged image is visible as yellow staining. The enlarged images are split screen enlargements of the boxed areas (from different frames of the Supplementary movie 1) with flotillin-1-GFP (green) at top of split and ephrinB2-mCherry (red) at bottom. Blue arrows indicate the same areas of co-localization in the membrane during movement. Scale bar represents 10 µm.