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Figure 1 Genetic and molecular characterization of the genomic region which contains the  TUB37C gene. In situ hybridization with a TUB37C probe maps this gene to bands 37C5–D1. The bars on the top depict the extent of the region deleted by some of the deficiencies which are available [for a detailed description of the deficiencies listed see Stathakis et al., (1995), and references quoted therein].  B1, B2 and B3 are overlapping inserts isolated from a Drosophila genomic DNA library made in -dash (C.Gonzalez, unpublished data). The arrow over the cDNA indicates the direction of transcription with respect to the restriction map shown above. We have not determined the orientation of the molecular map with respect to the polytene maps. P[TUB37C+ w+] is a 7.2 kb EcoRI fragment isolated from phage B3 which contains the TUB37C coding sequences flanked by 5 and 0.7 kb up- and downstream, respectively, cloned into the Drosophila transformation vector PW8. P[TUB37C- w+] is a derivative of P[TUB37C+ w+] generated by PCR, from which the coding region of the TUB37C gene has been deleted. The sequence at the bottom corresponds to the genomic sequence from the start (ATG) to the stop (TAA) codon. The two introns are represented by lower case letters.
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 | Figure 2 Developmental expression of the TUB37C polypeptide. Western blot hybridized with the Rb1011 antibody. Each line contains protein extracts from different tissues as indicated. (a) Egg chamber stages are as defined by King (1970). (b) Larval brains and salivary glands were obtained from third instar larvae. Adult testes were isolated from 2- to 10-day-old males. Balanced loading of total protein was controlled by scanning and quantitation following Ponceau staining of the membrane before incubation with the antibody. All the tracks contain similar amounts of protein except for embryos 2–7 h which contain  1/3 of the average and the adult testis track which was overloaded.
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Figure 3 Immunofluorescence analysis of the TUB37C polypeptide during embryogensis. (a), (c) and (d) were obtained following 3.7% formaldehyde fixation. The TUB37C polypeptide was visualized with the Rb1011 antibody (green/yellow) and DNA was stained with propidium iodide (red). (b) was obtained following methanol fixation. The TUB37C polypeptide is shown in orange, tubulin in green and DNA in blue. (a) Interphase nuclei from a syncytial blastoderm embryo. (b) High magnification of a single telophase. The TUB37C antigen co-localizes with the two MTOCs at each pole of the mitotic figure. A very prominent mid-body can be observed between the segregating chromatin masses. (c) High magnification of a single metaphase. Most of the TUB37C polypeptide is localized at the centrosomes, but significant amounts are also present over the spindle and scattered around the mitotic figure. (d) Area from the surface of an embryo during cell cycle 14, immediately after cellularization. Only mitotic cells show positive staining with the Rb1011 antibody.
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 | Figure 4 Developmental expression of the TUB37C polypeptide. Western blot hybridized with the Rb1011 antibody. The lines contain protein extracts from different tissues as indicated. Egg chamber stages are as defined by King (1970). Tissues were obtained from individuals which were wild-type (wt/wt), fs(2)TW11 homozygous (mut/mut), fs(2)TW11 hemizygous over Df(2L)VA23 (Df/mut) and fs(2)TW11 over the same deficiency plus the P[TUB37C+ w+] transgene that carries one copy of the TUB37C gene (P[+]/Df/mut). Balanced loading was controlled by scanning and quantification following Ponceau staining of the membrane before incubation with the antibody. The region of the scanned membrane containing the major bands present in ovarioles and early embryos is shown at the bottom of the figure.
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Figure 5 Meiotic figures in oocytes. (a), (b) and (c) are micrographs obtained from stage 14 oocytes. The panels in the bottom are drawings from oocytes at the same stage. (a) and the two drawings on the left top corner correspond to wild-type females. (b), (c) and the remaining drawings represent meiotic figures found in females which carried different heteroallelic combinations of mutants for the TUB37C gene.
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 | Figure 6 Immunofluorescence analysis of the meiotic spindle in fs(2)TW11 mutant oocytes. (a) Meiotic figure in a stage 14 oocyte from a wild-type female. (b–f) Meiotic figures found in oocytes from fs(2)TW11 homozygous females. Tubulin was visualized with the DM1a anti- -tubulin antibody (green) and DNA was stained with propidium iodide (red). All figures shown are projections made from 10–17 optical sections generated by confocal microscopy.
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Figure 7 Immunofluorescence analysis of embryos derived from fs(2)TW11 homozygous females. Distribution of DNA (a, b and c) and co-localization of DNA (red) and tubulin (green) (d, e and f) in 0- to 30-min-old embryos derived from wild-type (a and d) and fs(2)TW11 homozygous females (b, c, e and f). Tubulin was visualized with the DM1a anti--tubulin antibody and DNA was stained with DAPI.
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