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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 3. The ephrinB2 ectodomain is important for decreased ephrinB2 expression in the absence of flotillin-1.

Figure 3

(a) Deletion of the ephrinB2 ectodomain stabilizes its expression in the absence of flotillin-1. A schematic diagram of ephrinB2 and mutants is displayed, along with Western analysis of embryos injected with CoMO or F1aMO and wild-type (WT) ephrinB2, ephrinB2Δ60 or ephrinB2TM-Cyto RNAs. (b) ephrinB1 as well as ephrinB2 are associated with flotillin-1. Flotillin1a-HA or flotillin1b-HA RNA was injected with or without ephrinB1-Flag, ephrinB2-Flag, or ephrinB3-Flag RNA. Flotillin-1 was immunoprecipitated with anti-HA antibody, and immunoblotted with Flag antibody to detect ephrinBs. EphrinB1 and ephrinB2 were co-immunoprecipitated with Flotillin-1a and Flotillin-1b. Protein expression was confirmed in whole lysates by Western blotting with the indicated antibodies. (c) The ephrinB2 ectodomain makes ephrinB2 protein susceptible to decreased expression in the presence of the F1aMO. The domain structure of B1B2-HA and B2B1-HA chimeric proteins is displayed, along with Western analysis of embryos injected with F1aMO or CoMO and the indicated chimeric ephrinBs. Erk2 is used as a loading control.