Figure 5
From: Activated Ras alters lens and corneal development through induction of distinct downstream targets
Altered lens differentiation in Ras transgenic embryos. Immunohistochemistry (A-D, I, J, M-T) and in situ hybridizations (E-H, K, L) were performed on nontransgenic (NT) and Pax6-Ras transgenic embryos to detect expression of Pax6 (A, B), E-Cadherin (C, D, S, T), FoxE3 (E, F), Prox1 (G-J), Pitx3 (K, L), α-crystallin (M, N) and β-crystallin (O-T). In panels A-D, I, J and M-R, antigen antibody complexes are in red and in panels S and T, green and red. In all these panels nuclei are stained blue with DAPI. In situ hybridizations were performed using 35S-labeled riboprobes (E-H, K, L). Dark-field images were overlaid on respective bright-field images and silver grains were pseudocolored red. Upregulation of Prox1 (H, J) expression in a subset of Ras transgenic lens epithelial cells suggests initiation of early fiber differentiation. Co-localization of β-crystallin and E-Cadherin was seen in some of the Ras transgenic lens cells (T, arrows). The staining in the vitreous (panel C, asterisk) is due to anti-mouse secondary antibody binding to IgGs in the blood vessels. White arrows in panel G, H and I point to the lens equatorial region where fiber differentiation is initiated. Green arrowheads point to lens epithelial cells (G-J). Abbreviations; C, corneal epithelium; Cj, conjunctival epithelium; le, lens epithelium; lf, lens fibers; nr, neural retina. The scale bar (I) represents 100 μm in panels A-D, 25 μm in panels I, J, S, T and 50 μm in panels E-H, K-R.