Fig. 6: Exogenous expression of the human AQP0R33C mutation induces cortical cataract formation.

Embryonic chick lenses were injected with recombinant RCAS(A) expressing WT hAQP0 or the hAQP0R33C at E14 and E20. A Lysates of E14 chick lenses were prepared and immunoblotted with anti-AQP0 or anti-β-actin antibody. B E20 chick lenses were observed under a fluorescence microscope for AQP0WT-mcherry (red) and AQP0R33C-EGFP (green) expression in the whole lens, respectively (coronal orientations, upper panels). Frozen sections of the same lenses were prepared and fluorescence images were taken (middle panels for the whole lens, and lower panels for partial lens indicated with white box) and puncta numbers were quantified (bottom panel). For each group, n = 4. Scale bar: 400 µm for the upper two rows of panels. Scale bar: 50 µm for the bottom panel. C Opacity of E14 and E20 chick lenses expressing endogenous hAQP0 or hAQP0R33C mutations were shown by light transmission microscopy (upper panel) and quantified using the same method as described in Fig. 3B (bottom panel). For each group, n = 11. Scale bar: 400 µm. Sagittal paraffin sections of E14 (D) and E20 (E) chick lenses expressing WT hAQP0 or hAQP0R33C mutations were prepared and stained with H&E. Black arrowheads show empty and enlarged extracellular spaces. White arrowheads show abnormal nuclear distribution. Black asterisk points to a detachment region between epithelial and fiber cells. Scale bar, 50 µm. All data are presented as mean ± SEM. **P < 0.01, ****P < 0.0001. n = 3.