Figure 3.

The non-binding mutants Unc5^GU and GPC3^UG, Unc5-GPC3 interaction “in trans” is inhibited by “in cis” interactions

(A) A cell-based assay shows binding between mUnc5B (expressed on cells) and purified GPC3^core, but not GPC3^coreUG. Representative images.

(B) Quantification of cell-based assays. The Unc5-lingand FLRT2 (LRR domain, FLRT2^LRR) is also used.

(C) SPR binding data using Unc5B^ecto. The apparent K_D (K_Dcalc) was calculated using a 1:1 binding model and is indicative only.

(D) As (C), using rUnc5D^ecto.

(E) A cell-cell aggregation assay shows that GPC3 and Unc5D mediate cell adhesion in trans. Representative images.

(F) Quantification of cell-cell aggregation experiments. Empty vector controls: pCAGIC (red) and pCAGIG (green).

(G) Cell-cell aggregation assay using co-expression of Unc5D and GPC3 in cis. Representative images.

(H) Quantification of experiments using co-expression.

^∗∗∗p < 0.001; ^∗∗∗∗p < 0.0001, one-way ANOVA with Tukey’s post hoc tests. Scale bars represent 100 μm.

See also Figure S3.