Xenbase Image ID: 149041

Figure 6. The Transmembrane and the C2C Domains of E-Syt2 Are Both Required for Its Function(A) Schematic representation of the HA-E-Syt2 deletion mutants used.(B) Deletion of either the N-terminal or the C2C domain eliminates the dominant-negative effects of E-Syt2 gain of function on Xbra induction. Mutants were expressed and analyzed in AC induced with CA-FGFR1. Four independent experiments gave consistent results.(C) Only full-length E-Syt2 displays a dominant-negative gain of function effect on FGFR1 internalization. Each E-Syt2 mutant was analyzed in the receptor uptake assay in HEK293T cells as in Figures 5A and 5B. Upper panels show typical immunolocalization of initially membrane-labeled Myc-FGFR1 and of E-Syt2 mutants 20 min after bFGF addition. Scale bar, 10 μm. Lower panel shows statistical analysis of the data.(D) The N-terminal domain of E-Syt2 is required to mediate its interaction with FGFR1. E-Syt2 mutants were coexpressed with Myc-FGFR1 in HEK293T cells, and HA immunoprecipitates (I.P.) were analyzed by immunoblotting (I.B.).(E) The HA-δC2ABC mutant containing only the first 312 aa of E-Syt2 displays selective binding to activated FGFR1. The analysis in D was repeated for the mutant HA-δC2ABC, but cells were treated with either bFGF or with the inhibitor SU5402 before immunoprecipitation and the analysis of Myc-positive complexes.In (D) and (E), the asterisk indicates the unspecific detection of an endogenous protein by the HA antibody assay.See also Figure S6. Image published in: Jean S et al. (2010) Copyright © 2010. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

Return to previous page