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Figure 3


Fig. 3. Primary fiber cells fail to fully elongate to contact the anterior epithelium only in {alpha}8S50P/+ {alpha}3+/+ embryonic lenses even though typical gap junctions are still present. (A,B,D) Wild-type, {alpha}8-/- knockout and {alpha}8S50P/S50P {alpha}3+/+ embryonic lenses do not have any space between the epithelium and the underlying fiber cells at E15.5. (C) An E15.5 {alpha}8S50P/+ {alpha}3+/+ embryonic lens shows a large cystic lumen between posterior primary fiber cells and the anterior epithelium. All images were taken from Toluidine Blue-stained embryonic sections. (E,F) Triple-labeled images of {alpha}8 connexin (green), F-actin (red) and nuclei (blue) of frozen sections from wild-type (E) and {alpha}8S50P/+ {alpha}3+/+ (F) embryonic lenses, respectively. The punctate green fluorescent spots represent typical staining of {alpha}8 connexins in lens fiber cells. A large cystic lumen is present only in the {alpha}8S50P/+ {alpha}3+/+ embryonic lens in F. White arrowheads indicate the anterior ends of posterior fiber cells and a large white arrow indicates the anterior epithelium. (G) A high magnification image of the selected area in F shows punctate fluorescent spots (indicated by white arrows). (H) TEM shows bona fide gap junctions (indicated by white arrows) between posterior fiber cells of the E15.5 {alpha}8S50P/+ {alpha}3+/+ lens. Scale bars: 50 µm for A-D; 100 µm for E,F; 5 µm for G; 100 nm for H.