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Fig. S1. Alignment of full-length PDCD2 orthologs. Identical amino acids are shown in red, homologous amino acids in blue. K above the Drosophila sequence indicates E296K mutation in the Zfrp8M-1-1 mutant. Conserved regions and domains are outlined.
Fig. S2. Lymph gland phenotype of Zfrp8M-1-1 and northern blot of immune peptides in wild-type and mutant larvae. (A-D) The phenotypes of the lymph gland from wild-type (A) and Zfrp8M-1-1/ Zfrp8M-1-1 larvae (B). The lymph glands from Zfrp8M-1-1 homozygous and hemizygous animals develop melanotic masses in primary and secondary lobes (asterisks) during larval (B) and pupal stages (D). Wild-type (C) and Zfrp8M-1-1 (D) pupae. Primary lobes are marked with I, secondary with II. Scale bars: 70 μm in A,B. (E) Induction of the antibacterial peptide genes, Attacin, Drosomycin and Cecropin in wild-type and mutant third instar larvae. 15-20 larvae were bacterially challenged (Minakhina et al., 2003). After 2 and 6 hours, total RNA from ten larvae was extracted and the northern was hybridized with antibacterial peptide cDNAs and with RpS5 cDNA as an internal loading control. Unchallenged larvae were used as control.
Fig. S3. Immunostaining of centrosomes for CP309, Dgrip91 and γ-Tubulin of wild-type and mutant lymph gland cells. Confocal projections through one layer of cells of wild-type (A,C) and Df(2R)SM206 (B,D) lymph glands stained with anti-γ-Tubulin (A-D), anti-CP309 (A,B) and anti-Dgrip91 (C,D) to visualize centrosomes. DNA is stained with Hoechst33258. Scale bars: 5 μm.
Fig. S4. Immunostaining of apoptotic cells in wild-type and mutant lymph glands. Confocal images of wild-type (A), Zfrp8M-1-1 (B) and Df(2R)SM206 (C) lymph glands. Arrows indicate apoptotic cells with high level of Active Caspase 3 (red) and pyknotic nuclei. DNA is stained with Hoechst33258 (blue). Scale bars: 5 μm.
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