<i>src64</i> mutations and fertility of <i>src64^KO trans</i>-heterozygous females.

<p>Mutation is shown for each allele. + indicates the wild-type allele. For point mutations, the base pair change, the amino acid substitution and the protein domain containing the substitution are shown. Female fertility was assayed in females <i>trans</i>-heterozygous for the mutation and the <i>src64^KO</i> allele. Number of eggs laid by females over days 3–10 after introduction of males, is shown.</p>a<p>Altered nucleotide is underlined.</p>b<p>Wild-type amino acid is indicated, followed by position and mutant amino acid.</p

Ring canal growth defects in <i>src64</i> mutant egg chambers.

<p>Ring canals are stained with antibody to HTS. (A) Egg chambers. Ring canal diameters are reduced in the <i>src64^Δ17</i>, <i>src64^H402L</i>, <i>src64^D404N</i> and <i>src64^KO</i> mutants. Scale bar, 50 µm. (B) Typical ring canals ordered mean by diameter and oriented so diameter is presented along the vertical axis. Scale bar, 10 µm. (C) Ring canal diameters. A, does not differ from <i>+</i>; B, differs from <i>+</i> (P<0.05), but not other A alleles, except <i>src64^D204V</i> differs from <i>src64^R403C</i> (P<0.05); C, differs from <i>+</i> (P<0.001) and from A alleles (P<0.05) except <i>src64^P190L</i>, but not from group B, except <i>src64^D372N</i> differs from <i>src64^H402L</i> (P<0.01); D, does not differ from <i>src64^KO</i>, but differs from all other alleles (P<0.001). Error bars are SEM.</p

Nurse cell fusion defects in <i>src64</i> mutant egg chambers.

<p>Egg chambers are stained with phalloidin (green) and Hoechst (blue). Partial confocal projections are shown. Nurse cell fusion (arrows) occurs in <i>src64^Δ17</i>, <i>src64^H402L</i>, <i>src64^D404N</i> and <i>src64^KO</i> mutants. Nurse cell fusion does not occur in <i>src64^R403C</i> mutants. Scale bar, 50 µm.</p

Cellularization microfilament defects in <i>src64</i> mutant embryos.

<p>Embryos are stained with antibody to myosin II heavy chain. (A) Cross-sections showing the early cellularization front. Furrow canal depths are less uniform, producing a wavy and irregular cellularization front in <i>src64^Δ17</i>, <i>src64^H402L</i>, <i>src64^D404N</i> and <i>src64^KO</i> mutant embryos. Scale bar, 50 µm. (B) Grazing section projections of early cellularization stage embryos showing the microfilament network. Microfilament rings of <i>src64^Δ17</i>, <i>src64^H402L</i>, <i>src64^D404N</i> and <i>src64^KO</i> mutant embryos are irregular. Scale bar, 10 µm. (C) Grazing section projections of late cellularization stage embryos showing microfilament rings. Microfilament rings of <i>src64^Δ17</i>, <i>src64^H402L</i>, <i>src64^D404N</i> and <i>src64^KO</i> mutant embryos are irregular and less constricted than the microfilament rings of wild-type embryos and <i>src64^R403C</i> mutant embryos. Scale bar, 10 µm. (D) Microfilament ring circularity during early cellularization. A, does not differ from <i>+</i>; AB, differs from <i>+</i> and <i>src64^Δ17</i>; B, does not differ from <i>src64^Δ17</i> allele but differs from <i>+</i> (P<0.01); C, does not differ from <i>src64^KO</i> but differs from <i>+</i> (P<0.001) and <i>src64^Δ17</i> (P<0.001). Error bars are SEM. (E) Microfilament ring circularity during late cellularization. A, does not differ from <i>+</i>; B, does not differ from <i>src64^Δ17</i> but differs from <i>+</i> (P<0.05); C, does not differ from <i>src64^KO</i> but differs from <i>+</i> (P<0.001) and <i>src64^Δ17</i> (P<0.001). Error bars are SEM.</p

Cytoskeletal defects of <i>src64^D404N/src64^KO trans</i>-heterozygotes.

<p>(A) <i>src64^D404N/+</i>, <i>src64^D404N</i> and <i>src64^D404N/src64^KO</i> egg chambers stained with antibody to HTS. Ring canal diameters are not reduced in <i>src64^D404N/+</i> egg chambers but are reduced in <i>src64^D404N/src64^KO</i> egg chambers. Inset: ring canal of approximately mean diameter, reoriented so that diameter is projected along the vertical axis. Scale bar, 50 µm. (B) Early cellularization stage embryos laid by <i>src64^KO</i>, <i>src64^D404N</i> and <i>src64^D404N/src64^KO</i> mothers. Embryos show non-uniform cellularization fronts. Scale bar, 100 µm. (C) Grazing section projections of early cellularization stage embryos laid by <i>src64^KO</i>, <i>src64^D404N</i> and <i>src64^D404N/src64^KO</i> mothers showing microfilament networks. Microfilament ring defects of maternally <i>trans</i>-heterozygous <i>src64^D404N/src64^KO</i> embryos are similar to those of <i>src64^KO</i> and <i>src64^D404N</i> embryos. Embryos are stained with antibody to myosin II heavy chain. Scale bar, 10 µm. (D) Ring canal diameters. <i>src64^D404N</i> and <i>src64^D404N/src64^KO</i> do not differ (p = 0.35). Error bars are SEM. (E) Microfilament ring circularity during early cellularization. <i>src64^D404N</i> and <i>src64^D404N/src64^KO</i> do not differ (p = 0.52). Error bars are SEM.</p

Tyrosine kinase activities of <i>src64</i> HRD mutants.

<p>Phosphotransfer was measured in cellularization-stage embryo extracts. Src64-dependent phosphotransfer to a peptide substrate is shown. <i>src64^H402L</i>, <i>src64^D404N</i> and <i>src64^KO</i> differ from wild type (p<0.01). Wild type, <i>src64^H402L</i> and <i>src64^R403C</i> differ from <i>src64^KO</i> (p<0.01). Error bars are SEM.</p