Figure 1
Deleting Ate1 in the brain and the retina. (A) Strategy for generating Ate1 conditional knockout (KO) mice. (B) Western blots using ATE1 antibodies confirm near-absence of ATE1 in the brain (left) and significant reduction of ATE1 in the retina (right) of Ate1 KO mice. Western blots were performed on brain and retina lysates from littermate pairs of wild type (WT) and KO mice. For the retina blot, WT and KO pair were on the same blot but not on the adjacent lanes. (C) Immunohistochemistry of central retina sections stained with anti-ATE1 (green) in WT (top) and KO (bottom), together with the G-protein G\(\alpha _{o}\) (red, the marker for the ON bipolar cells) and DAPI (blue, the marker for the nuclei). The right-hand images show a 3.4x magnification of the regions of the retina shown on the left. The \(+\) signs mark two representative WT and KO ON-bipolar cells, respectively. Shown here are original images of KO and WT processed at the same time with the same settings. See Fig. S1A for the corresponding images with background subtraction and/or scaling. The WT retina has clear ATE1 staining in the ONL, OPL, INL, IPL, GCL, and ON-bipolar cells; this ATE1 staining is greatly reduced in KO except bright spots that stained blood vessels (marked ‘bv’) and a few cells in INL near the border with IPL. See Fig. S1B for another example. IPL/OPL: inner/outer plexiform layer. INL/ONL: inner/outer nuclear layer. GCL: ganglion cell layer. (D) and (E), Quantification of ATE1 and G\(\alpha _{o}\) levels in different retinal layers. The error bars represent SEM and the p-values are from paired Student’s t-test on 6 pairs of retinal sections from WT and KO littermates. Data points from each pair are plotted with a unique symbol.
