Fig. S3. Loss of PI3K-C2? leads to cataract in zebrafish and human, and increased senescence in the mouse lens. (A) Adult zebrafish were anesthetized with tricaine and examined. Coaxial illumination using a Leica M841 surgical microscope was used to visualize lenticular defects. Optical sectioning by changing the z-axis focus was used to aid in the identification of cataracts. All zebrafish examined for cataracts were offspring of a cross between pik3c2asa10124/+ and pik3c2asa12328/+ fish. (B) Representative images of posterior lenticonus cataract in one of the PIK3C2A-null patients. Arrows indicate opacities in lens. (C) Representative images of eyes and their H&E-stained sections derived from a wild-type and a Pik3c2ahypo/hypo mouse 30 days after birth. (D) Immunohistochemistry using anti-KI-67 antibody in eye sections from wild-type and Pik3c2ahypo/hypo mice at 30 days after born and quantification of percentage of KI-67 positive nuclei in wild-type and Pik3c2ahypo/hypo mice. (E) Immunohistochemistry using anti-p16INK4A antibody in eye 5 sections as in (C) from wild-type and Pik3c2ahypo/hypo mice at 30 days after birth. If not previously specified, all results are shown as mean or representative picture of at least three independent experiments ± SEM (*P<0.05; **P<0.01; ***P<0.001).
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