Fig. S2

Lens epithelial cells enter fiber differentiation without passing through the equator in rw341 mutants

(A, B) Labeling of wild-type (A) and rw341 mutant (B) lenses with anti-Prox1 antibody. Nuclei were counterstained with TOPRO3. Right panels indicate higher magnification of lens epithelium area shown by squares in the left panels. In rw341 mutants at 3dpf, some cells express Prox1 in monlayered lens epithelium (B, arrowheads) . Prox1-positive cells increase and the monolayer of lens epithelium is disrupted at 4 dpf. (C, D) Prox1 (C) and AQP0 (D) expression in 9 dpf wild-type and rw341 mutant lenses counter-labeled with anti- PCNA antibody. In rw341 mutant lenses, aggregated anterior lens cells are observed close to the transparent lens fiber core. Only a few cells express Prox1 (C arrowheads), and most cells express AQP0 (D, asterisk). AQP0- positive cells are still nucleated, suggesting that denucleation process may be compromised. (E) Developmental profile of lens phenotypes in rw341 mutants. rw341 mutant lens epithelial cells start to express Prox1 at 3 dpf, and then Prox1 expression switches to AQP0 expression after 5 dpf. Most cells express AQP0 to form lentoid at 9 dpf. Scale: 20 μm (A, B). Development: doi:10.1242/dev.170282: Supplementary information Development • Supplementary information