1. Volume 20, Issue 6, Pages 821-831 (December 2005)
The Putative Drosophila Transcription Factor Woc Is Required to Prevent Telomeric Fusions 
Grazia D. Raffa, Giovanni Cenci, Giorgia Siriaco, Michael L. Goldberg, Maurizio Gatti 
Molecular Cell 
Volume 20, Issue 6, Pages (December 2005)
DOI: /j.molcel
Copyright © 2005 Elsevier Inc. Terms and Conditions

2. Figure 1 Examples of Telomeric Fusions Found in woc Mutants
(A) Control (Oregon R) male metaphase.
(B) Metaphase showing a tricentric chromosome generated by an XL-2 DTA (asterisk) and a 2-2 STA (arrowhead).
(C) Metaphase containing three dicentric chromosomes generated by XR-XR (arrowhead), 2-3 (asterisk), and 4-4 (arrow) DTAs.
(D) Metaphase with a dicentric ring involving the two X chromosomes (arrowhead) and a linear dicentric generated by a 2-2 DTA (asterisk).
(E) Metaphase showing two linear dicentrics generated by 2-2 (asterisk) and 3-3 (arrow) DTAs.
(F) Aneuploid metaphase containing a tetracentric and a dicentric chromosome, generated by (arrow) and 2-4 (arrowhead) DTAs, respectively.
Molecular Cell  , DOI: ( /j.molcel )
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3. Figure 2 Characterization of woc Mutants and Their Protein Products
(A) Structure of the Woc protein and localization of the mutations. The wt Woc protein contains 1688 amino acids and is characterized by eight Zn finger (filled bars) and two AT-hook (empty lozenges) motifs. The wocB111, woc468, and woc251 alleles contain premature stop codons (positions shown in brackets), resulting in truncated proteins of 553, 754, and 1474 amino acids, respectively. In the protein product of the woc964 allele, an aspartate residue at position 1464 is converted into a tyrosine.
(B) Western blots showing that the anti-Woc230–626 antibody recognizes an ∼187 kDa band in larval brain extracts from both wt and woc964 mutants; this band is absent in brain extracts from wocB111, woc468, and woc251 mutants. α-tubulin was used as a loading control (LC).
(C) Localization of the Woc protein on prometaphase chromosomes from a wt larval brain. Chromosomes were stained with DAPI and with the anti-Woc230–626 antibody. The panels at the bottom show a particularly elongated Y chromosome displaying its typical subdivision into differently stained regions (indicated by numbers). Note that Woc is enriched at the tip of XR, the fourth chromosomes, and several regions of the Y chromosome. See text for further explanation.
Molecular Cell  , DOI: ( /j.molcel )
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4. Figure 3 Localization of Woc on Polytene Chromosomes
(A) Wt polytene chromosomes immunostained for Woc showing hundreds of Woc-positive regions.
(B) Staining of wt chromosomes for DNA and Woc reveals that most Woc signals are localized in the interbands, as evident in the split image.
(C) Double immunostaining of wt chromosomes for Woc and HP1. In the merged image the Woc (red) and HP1 (green) signals at telomeres overlap (yellow), indicating colocalization of these proteins.
(D) Examples of polytene chromosomes (2L distal regions) from wt and woc964 mutants. Although mutant chomosomes display an overall Woc staining pattern comparable to wt, they are usually devoid of Woc signals at telomeres (arrows).
Molecular Cell  , DOI: ( /j.molcel )
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5. Figure 4
Woc Colocalizes with Initiating Forms of Pol II but Fails to Associate with Heat-Shock-Induced Puffs
(A) Woc (red) extensively colocalizes with both Pol IIa and Pol IIoser5, but not with Pol IIoser2 (all Pol II forms are in green).
(B) Woc and Pol II distributions in polytene chromosomes exposed to 1 hr heat shock. The enlarged images in the right panels clearly show that Woc fails to accumulate at heat-shock-induced puffs.
Molecular Cell  , DOI: ( /j.molcel )
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6. Figure 5
Functional Relationships between woc, Su(var)205, cav, tefu/atm, and rad50
(A) Wt and wocB111 mitotic chromosomes immunostained for HOAP (red) and counterstained with DAPI.
(B) Wt and wocB111 polytene chromosomes (2R distal regions) immunostained for both HP1 and HOAP; HP1 staining is shown in black and white, whereas HOAP staining (red) is merged with DAPI staining (black and white). Note that the telomeres of mutant chromosomes accumulate normal amounts of HOAP and HP1.
(C) Distal regions of polytene X chromosomes immunostained for Woc. Chromosomes from cav/cav, Su(var)20504/Su(var)20505, rad50Δ5.1/rad50Δ5-1, and atm6/atm6 mutant larvae display telomeric Woc signals comparable to wt.
Molecular Cell  , DOI: ( /j.molcel )
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7. Figure 6
Mutations in woc Do Not Dominantly Affect Drosophila Telomere Length
(A) Quantification of HeT-A and TART sequences by slot blotting. Slot blots from wt and woc mutant strains (251, 468, and 964) were probed with a mixture of HeT-A and TART probes or with total genomic DNA (“LC,” loading control); a Tel-bearing strain was used as a positive control. The histogram represents the ratios of the HeT-A and TART signal relative to the corresponding loading control; the ratio for the wt sample is equal to one.
(B) Quantification of HeT-A and TART sequences by FISH to polytene chromosomes of wt and woc/TM6C strains. Each panel shows examples of the distal portions of the XL (top) and 3L (bottom) arms hybridized with a mixture of HeT-A and TART sequences. The histogram shows the mean intensities of the FISH signals (± SE; n > 15 for all samples) relative to the wt mean value.
Molecular Cell  , DOI: ( /j.molcel )
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