Figure 2. ITR-1 Expression during Ventral Enclosure. Images show representative wild-type and itr-1
) embryos expressing AJM-1::GFP; the embryos were stained for ITR-1 (antibody 263, a gift from K. Mikoshiba).(A, D, G, and J) AJM-1::GFP; (B, E, H, and K) anti-ITR immunostaining; (C, F, I, and L) merge.(M) Western blot analysis of ITR (left) and SMG-3 (right) (a gift from P. Anderson) as a loading control.(A)-(F) are ventral views; (G)-(L) are dorsal views. All embryos are shown with anterior to the left and posterior to the right.(A-C) Wild-type embryo undergoing enclosure.([B], inset) 2.5 magnification of a portion of two ventral epidermal cells from another embryo immunostained for ITR-1; the magnified region shows reticular staining.(C) ITR-1 staining is evident at epidermal cell borders, as shown by overlapping signal (arrow).(D-F) itr-1
) embryo undergoing enclosure. Leading cells are starting to retract (arrowhead). There is no ITR-1 staining evident at epidermal cell membranes.(G-I) Wild-type embryo undergoing dorsal intercalation.(H) ITR-1 localizes near the plasma membrane (arrow) and in a reticular pattern.(J-L) itr-1
) embryo undergoing dorsal intercalation. Dorsal cells have not intercalated (asterisks).(K) Where dorsal cells have not intercalated properly, ITR-1 staining appears more diffuse, and membrane staining is not visible. The scale bar represents 10 μm.