Figure 1. KCC2 and NKCC1 expression in a 30-day-old mouse lens. A: KCC2 mAb immunofluorescence detection (400X magnification) in mouse lens corresponds to the arrowhead in panel D. The lens perimeter is at the left. B: Anti-KCC2b (400X magnification; inset plus ~ 2X photomagnification). KCC2 is detected at the fiber cell borders in the interior lens fiber cells in the inset photo. Mouse lens fiber cells are about 7 μm in their longest dimension. C: Anti-KCC2b (600×, 3X photomagnification) identifies expression in cortical fiber cells as they terminally differentiate to mature fiber cells (arrowheads indicate the sites of the unstained cell nuclei). D: KCC2 is detected in interior fiber cells in the mouse lens (100X). The asterisk identifies the corresponding position in panel E and identifies KCC2 expression in the lens interior. The arrow at the lower left indicates the lens perimeter. E: NKCC1 mAb (monoclonal Ab) detection of cortical fiber cells with little detection in the interior lens nucleus. F: Lower exposure photo showing anti-NKCC1 polyclonal antibody detection restricted to lens cortical fibers in the mouse lens (200X, original magnification). G: KCC2b lower exposure detects protein in the cortical fiber cells. H: No primary antibody control. I: KCC2 detected at a site corresponding to the arrowhead in panel D. J: Actin detected in the same view as in panel H. K: I, J merged images. L: Immunoblot identifies KCC2b expression in the total mouse lens protein samples.