Fig. S3

Expression of Pax6, ZO-1, Pcna, Prox1, foxe3 and αA-crystallin mRNA in wild-type and vov mutant embryos. (A,B) Expression of Pax6 (green) and ZO-1 (red) in wild-type (A) and vov mutant (B) lens at 72 hpf. (A′,B′) High magnification of anterior lens epithelium in A and B. (A′′,′′2) The same images of A and B shown through a red channel (ZO-1). In the vov mutant, ZO-1 signals underneath anterior lens epithelium are weaker than in wild type and sometimes discontinuous (B′′). (C,D) Expression of Pcna (green) and Prox1 (red) in wild-type (C) and vov mutant (D) lens at 72 hpf. In wild type, Pcna (green) is expressed strongly in the anterior lens epithelium and also weakly in four or five cells that pass over the region adjacent to the retinal CMZ (arrowheads, C); the latter are labeled with Prox1 (red). In the vov mutant, Pcna (green) is not only expressed in the anterior lens epithelium but also in cells located at the posterior edge of the lens sphere. Some Pcna-expressing cells strongly express Prox1 (red) at the posterior lens edge in the vov mutant (arrowheads, D). (C′) High magnification of the posterior lens region of C. (C′′,C′′′) The same image as C′ through a green (Pcna) or red (Prox1) channel. The Pcna and Prox1 double-positive cells seem to be early differentiating lens fiber cells. Asterisks indicate cells of intraocular blood vessels. (D′) High magnification of the posterior lens region in D. (D′′,D′′′) The same image as D2 through a green (Pcna) or red (Prox1) channel. Most posterior Pcna-positive cells are labeled with Prox1 (red, arrowheads), suggesting that posterior-positioned Pcna cells in the vov mutant are early differentiating lens fiber cells. (E,F) foxe3 mRNA expression of wild-type (E) and the vov mutant (F) lens at 72 hpf. foxe3 mRNA is expressed in the lens epithelium in wild type and the vov mutant. (G,H) Expression of crystallin αA mRNA in 72 hpf wild-type (N) and vov mutant (O) lens. crystallin αA mRNA is strongly expressed in differentiating lens fiber cells in wild type and the vov mutant. Scale bars: 50 µm, except 10 µm in A′,A′′,B′,B′′.