Figure 4
Water-soluble αA-crystallin in R49C neo knock-in lenses. Protein solubility was assessed in lenses with different nuclear cataract severity ranging from stage 1 to 3 as determined by slit lamp analysis. (A) WT lenses had the highest protein solubility by immunoblotting with an antibody to α-crystallin. The wild type is clear lens (stage 0). A decrease in soluble protein levels was seen in the WT/R49Cneo lenses. R49Cneo/R49Cneo lenses had a significantly lower water solubility. Proteins were overloaded so that even a small amount of protein could be visualized. Under these conditions, cross-reactivity of the antibody with other crystallins was observed. (B) The decrease in levels of soluble proteins was determined by densitometric scanning of immunoblots. The data represent an average of three independent experiments with approximately 3-month-old mouse lenses. Three pairs of lenses were analyzed for wild type and R49Cneo/R49Cneo homozygous mice. Two pairs of lenses were analyzed for WT/R49Cneo heterozygous mice.