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Fig. 4. Crystallin expression and its regulation in Sip1-defective lenses. (A-E) Normal lenses and (F-J) Sip1-defective lenses, subjected to immunostaining (A-C,F-H) and in situ hybridization (D,E,I,J). (A,F) At E12.5, {alpha}-crystallins are expressed throughout the lens under either normal (A) or Sip1-defective (F) conditions. ßA-crystallins are normally expressed widely in nonepithelial domains through immature to mature lens fibers (B), but in Sip1-defective lens, its expression was confined to the innermost domain (G). {gamma}-Crystallin expression marking mature lens fiber cells in normal lens (C) is totally missing in Sip1-defective lens (H), clearly indicating a defect in lens fiber maturation. Broken lines indicate the boundaries of the bow region. However, two known key transcription factor genes for {gamma}-crystallin regulation, Sox1 (Kamachi et al., 1995; Nishiguchi et al., 1998) and Maf (Kawauchi et al., 1999; Kim et al., 1999; Ring et al., 2000), are expressed in normal (D,E) and Sip1-defective (I,J) lenses. Scale bars: 100 µm.