Journal: Molecular Vision
Article Title: KCC2 expression supersedes NKCC1 in mature fiber cells in mouse and rabbit lenses
doi:
Figure Lengend Snippet: 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.
Article Snippet: Antibodies were obtained from the following sources: KCC2, NKCC1, and GABBR1 mAbs from Neuromab/Antibodies Inc. (Davis, CA), anti-NKCC1 from Alamone Labs (Jerusalem, Israel), and anti-KCC2b, anti-GABAAβ3, and STEP mAb from Santa Cruz Biotechnologies.
Techniques: Expressing, Immunofluorescence, Western Blot