Cataracts and Microphthalmia Caused by a Gja8 Mutation in Extracellular Loop 2
Figure 3
Biochemical and immunological characterization of Cx50 and Cx46 proteins.
(A) Total lens homogenates, prepared from Gja8R205G/R205G (a), Gja8R205G/− (b) and Gja8−/− (c) littermates at one week of age, were examined by a Coomassie-blue stained gel (left panel) and by western blot using an anti-Cx50 antibody and an anti-Cx46 antibody (right panels). Arrowheads indicate phosphorylated proteins while the arrow indicates non-phosphorylated proteins. Compared to Gja8−/− lenses, mutant Gja8R205G/R205G lenses showed decreased phosphorylated Cx46 proteins. (B) Immunostaining results of lens frozen sections showed typical punctate signals of Cx50 (red) and Cx46 (green) connexin proteins in wild-type (Gja8+/+Gja3+/+) sample, but only diffuse tiny spots of Cx50 and Cx46 could be seen in Gja8R205G homozygous mutant (Gja8R205G/R205G Gja3+/+) sample. Diffuse tiny spots of Cx50 were observed in Gja8R205G Gja3 double mutant (Gja8R205G/R205G Gja3−/−) sample. These lens samples were prepared from 1-week-old mice. Scale bar, 20 µm.