Fig. S2

Fig. S2 crystallin insolubility and rescue in cloche^s5. (A) Western blot: total cloche^m39 and cloche^s5 embryo extracts were analyzed by western blotting with anti-αA-crystallin antibody. (B) Zebrafish embryo (2-4 dpf) mRNA levels were analyzed by semi-quantitative RT-PCR. Wild-type versus cloche^s5 embryo gene expression is shown for αA-crystallin and γ-crystallin. ef1a was used as a loading control. (C) αA-crystallin overexpression prevents γ-crystallin insolubility. Zebrafish embryos (2.5 dpf) were lysed detergent-free in 20 mM Tris-HCl, pH 7.5, and centrifuged. Soluble (top) and insoluble (bottom) fractions were separated, and were each analyzed by western blot with anti-γ-crystallin antibody. Two independent experiments using cloche^s5 were performed.