FIG. 1.

Molecular analysis of mutant Psc proteins. (A) Sequence analysis. Diagrams depict wild-type Psc (black), proteins of the moderate alleles (red), strong alleles (green), and engineered deletion constructs (white). Products of strong alleles that are likely associated with expression defects are shown in light green, and asterisks indicate locations of missense amino acid changes. The homology region (gray shading), Ring finger (dark stripes), and HTHTH motifs (light stripes) are indicated. (B) Western analysis of animals heterozygous for mutant Psc alleles. (C) Western analysis of embryos homozygous for Psc¹⁴⁴⁵ or Psc^e23. Equal protein loading was confirmed by Amido Black staining (for Psc¹⁴⁴⁵) or by Western blotting using an antibody to TATA binding protein (TBP) (for Psc^e23; Santa Cruz). (D and E) Recombinant Psc proteins corresponding to mutant alleles (∼400 ng) (D) or engineered deletions (∼500 ng) (E) were separated on 4 to 15%-gradient SDS-PAGE gels and stained with colloidal Coomassie. An asterisk (D and E) indicates contaminating heat shock protein 70 (Hsp70).