Figure 3.

N-cadherin Variants Have Impaired Cell-Cell Adhesion and Reduced trans Adhesion to N-cadherin-WT

(A) Schematic representation of the cell-aggregation assay. L cells were transfected with empty GFP, N-cadherin-WT, and N-cadherin variants. Aggregation was then performed for 30 min in the presence of calcium.

(B) Bar graph of the mean ± SE of the index of aggregation for N-cadherin-WT (black) and N-cadherin variants (gray) (n = 5 experiments). Cell aggregation was evaluated after 30 min of aggregation (T30) by the index (N₀-N₃₀)/N₀, where N₀ is number of GFP-positive cells or aggregates at T0, and N₃₀ is the number of GFP-positive cells or aggregates at T30. The N-cadherin-GFP variants have a lower index of aggregation than N-cadherin-WT-GFP cells do. One-way ANOVA post-tests were performed for each variant versus N-cadherin-WT (^∗∗∗∗p ≤ 0.0001; ^∗∗∗p ≤ 0.001; ^∗∗p ≤ 0.01; ^∗p ≤ 0.05).

(C) Schematic representation of the mixed-cell aggregation assay evaluating trans interaction with N-cadherin-WT. L cells were transfected with empty GFP, N-cadherin-WT, and N-cadherin variants or with N-cadherin-WT-Myc+mCherry. Cells were then mixed in a 1:1 ratio, and aggregation was performed for 30 min in the presence of calcium.

(D) Bar graph of the mean ± SE of the fraction of mixed aggregates for N-cadherin-WT (black) and N-cadherin (gray) variants (n = 3 experiments). The fraction of mixed aggregates was evaluated after 30 min of aggregation (T30) by the following index: number of mixed aggregates/number of total aggregates. Cells expressing N-cadherin WT-myc+mCherry form mixed aggregates with cells expressing N-cadherin-WT-GFP, but they form fewer mixed aggregates with cells expressing the N-cadherin variants. One-way ANOVA post-test were performed for each variant versus N-cadherin-WT (^∗∗∗p ≤ 0.001; ^∗p ≤ 0.05).