Fig 2: Effects of the GEFS+ mutations on the cell-cell adhesion.(a) Mapping of the GEFS+ mutations on the ß1 model structure. Side view (upper) and top view (lower). The model was generated by the superimposition of the monomer model (built by the program MODELLER37) onto the ß4 structure in Fig. 2a. (b) Close-up view of the C?–D loop interaction. The hydrogen bonds are indicated by light blue lines. (c) Immunofluorescence staining of Neuro2A cells stably expressing the WT and GEFS+ mutants of ß1. The ß1 proteins were immunostained with anti-ß1ex against the N-terminal residues 70–98 of the ß1 extracellular domain (Abcam ab107370), and visualized by Alexa 647 (magenta). Nuclei were stained with Hoechst 33342 (cyan). (d) Aggregation of Neuro2A cells stably expressing the WT and GEFS+ mutants of ß1. Results are expressed as N30/N0, where N30 is the number of particles after a 30 min incubation, and N0 is the number of particles at the start. ***P < 0.001 (one-way ANOVA, means ± S.E.M, n = 48 from three independent experiments).