FIGURE 3:

The extracellular domain of Tril is required to retain it at the plasma membrane while the intracellular domain is required for internalization. (A–D‴) RNAs encoding wild-type or deletion mutant forms of Tri-HA (illustrated to the left of each panel) and membrane-localized RFP (memRFP) were injected into a single animal pole blastomere of four-cell embryos. Ectoderm was explanted from 5–10 embryos in each group at stage 11 and immunostained for HA and RFP. (A–D″) Projections viewed from the xy-axis. (A‴–D‴) Projections viewed from the z-axis. Results were replicated in three independent experiments. (E) CHO cells transfected with GFP (negative control), cadherin6-GFP (Cad6-GFP) (positive control), or Tril-GFP were tested for adhesion. Only cells expressing cadherin6 formed aggregates. The aggregation index was calculated by dividing the total GFP fluorescence in cell aggregates by the total GFP fluorescence in the well. Mean ± SD are shown, n = 3, ***p < 0.01 by two-tailed t test.