Supplementary Figure 1. Measurement of the Truncated Mip130 Protein Levels in Mip130 Mutants
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Supplementary Figure 1. Measurement of the truncated Mip130 protein levels in mip130 mutants.
Adult mutant males of the indicated genotype were homogenized in sample buffer and one, two, and four microliters of extract examined for Mip130 protein levels as indicated at the right by immunoblot analysis. Tubulin was used as a loading control.
Chemiluminescence levels were quantitated using a Typhoon (Amersham) and the
Mip130* signal relative to the Tubulin signal was: 17% for mip1301-723+1-36, 22% for mip1301-723, 19% for Dm-myb mip1301-723+1-36, and 22% for Dm-myb mip1301-723.
Supplementary Figure 2. FACS analysis of mip130 mutant follicle cell nuclei.
Nuclei isolated from ovaries of the indicated genotypes were stained with propidium iodide (PI) and subjected to FACS analysis in order to determine the DNA content of the follicle cells. Mitotically active follicle cell nuclei give rise to the 2C peak and the ensuing three endocycles give rise to the 4C, 8C, and 16C peaks. The small 32C peak is always ovserved in wild type preparations and is derived from contaminating nurse cell nuclei as described (Cayirlioglu et al. 2001). (A and B) Nuclei isolated from heterozygous mip130 mutant ovaries as indicated. (C and D) Nuclei isolated from homozygous mip130 mutant ovaries as indicated. Neither the null mutant (D) nor mip1301-723+1-36 (C) showed an increase in the 32C peak when compared to heterozygous mutant sisters indicating that an additional complete endocycle did not occur in these mutants.