Fig. 8: Protein aggregation results from human crystalline mutations exogenously expressed in the embryonic chick lens.
From: Development of a potent embryonic chick lens model for studying congenital cataracts in vivo
Embryonic chick lenses without injection (A) or injected with recombinant RCAS(A) expressing WT human αA-crystalline (hCRYAA) (B), or one of two mutations, hCRYAAR12C (C), and hCRYAAR54C (D). Lysate of E20 lenses was prepared and fractionated on a linear gradient of 4–20% sucrose. Each fraction was subjected to SDS/PAGE and western-blotted with an anti-αA-crystallin antibody (upper panels). The sedimentation coefficients were calculated and the band intensity of αA-crystalline in each gradient fraction on western blots was quantified by Image J. and normalized as a percentage of the fraction containing the highest level of hCRYAA or mutations, respectively (bottom panels).
