Figure 5.

The development of polarization of tropomyosin isoforms. (A-L) immunofluorescence confocal microscopy images of T84 cells stained for tropomyosin isoforms at various time points after seeding. All images are in the vertical (xz) plane. At each time point, the images on the left and in the center are of the same costained cells. On the left (A, D, G, and J) the 311 antibody (Tm 3, 6) staining is shown. In the center (B, E, H, and K) the αf9d antibody (Tm 3, 5a, 5b, 6) staining is shown. The cells on the right (C, F, I, L) are stained with CG3 antibody (TmNM1–11). (A, B, and C) 10 min; (D, E, and F) 1 h; (G, H, and I) 2 h; (J, K, and L) 24 h. The arrows indicates a T84 cell in suspension with circumferential staining. Bar, 10 μm. (M and N) Total protein and specific tropomyosin isoform expression during T84 cell monolayer development. Protein was extracted from T84 cells 1, 2, 4, and 24 h and 7 d after seeding. (M) Gel stained with Coomassie blue showing total protein. (N) Western blot immunoblotted with αf9d antibody (Tm 3, 5a, 5b, 6).