alnz24 is a null mutation that regulates NC formation and encodes RNA polymerase II-associated factor 1 (Paf1). (A-E) 36?hpf wild-type and aln mutant embryos. Wild-type embryos (A) have pigmented melanophores lacking in embryos harboring combinations of aln mutant alleles: (B) homozygous for the alnz12 deletion mutation, (C) transheterozygous for the alnz12 and c4 deletions, (D) transheterozygous for the alnz12 deletion and alnz24 ENU-induced mutations, and (E) homozygous for the alnz24 mutation. (F,G) crestin expression detected by whole-mount in situ hybridization in 24?hpf wild-type sibling (F) and alnz24 mutant (G) embryos, indicating a complete absence of migrating trunk NC in alnz24. Arrowheads indicate premigratory and migrating trunk NC. A-G are lateral views with anterior to the left. (H) Paf1 protein is not detected by immunoblot analysis in 36?hpf alnz12 or alnz24 mutant embryos, but is readily detected in wild-type control siblings. Immunoblot detection of pMAPK serves as a protein-loading control.
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