Ectopic lens fiber differentiation depends on TGFβ signaling. (A) 5 dpf wild-type and rw341 mutant lenses treated with DMSO and SB505124. In DMSO-treated rw341 mutants, lens epithelial cells form multilayers (asterisk). SB505124 treatment rescues multilayer phenotypes in rw341 mutants (arrowheads). (B) Percentage of transparent lens fiber area in wild type and rw341 mutants treated with DMSO and SB505124. SB505124 treatment significantly increases transparent fiber area size in rw341 mutants, although the recovery does not reach the wild-type level. (C) Prox1 (green) and PCNA (magenta) expression in 5 dpf rw341 mutant lenses treated with DMSO and SB505124. Lower panels indicate higher magnification of lens epithelia (le). lf, lens fiber. Upper right and lower panels show only the green channel. In DMSO-treated rw341 mutants, most cells express Prox1 in multilayered lens epithelium (le). In BS505124-treated rw341 mutants, the number of Prox1-positive cells is markedly reduced (arrowheads). Asterisks indicate the Prox1-negative lens epithelial area. (D) AQP0 (green) and PCNA (magenta) expression in 5 dpf rw341 mutant lenses treated with DMSO and SB505124. Lower panels indicate higher magnification of lens epithelia. Upper right and lower panels show only the green channel. In DMSO-treated rw341 mutants, AQP0-positive lentoid-like structures are observed (arrowheads). In SB505124-treated rw341 mutants, only one PCNA-positive cell expresses AQP0 (arrowhead), but lentoid-like structure is not observed (asterisks). (E) The number of Prox1-positive cells in anterior lens epithelium per lens section. (F) Percentage of Prox1-positive cells relative to the total number of lens epithelial cells. (B,E,F) Data are mean±s.d. (B) Two-way ANOVA. (E,F) Student's t-test: *P<0.05, **P<0.01, ***P<0.005. Scale bars: 20 µm.
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