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1.
FIG. 1

FIG. 1. From: Meiotic Telomere Distribution and Sertoli Cell Nuclear Architecture Are Altered in Atm- and Atm-p53-Deficient Mice.

PNA telo-FISH to testis suspension nuclei of Atm-p53-deficient mice. (a and b) Telomeres (whitish) are scattered throughout premeiotic, somatic nuclei (gray), as seen at the maximum nuclear diameter. (c and d) Top view of spermatocyte I nuclei which display telomeres clustered at a limited sector of the nuclear periphery (indicative of leptotene-zygotene transition). DNA was counterstained with DAPI (gray).

Harry Scherthan, et al. Mol Cell Biol. 2000 October;20(20):7773-7783.
2.
FIG. 3

FIG. 3. From: Meiotic Telomere Distribution and Sertoli Cell Nuclear Architecture Are Altered in Atm- and Atm-p53-Deficient Mice.

H1t (red) and anti-SCP3 (green) immunostaining of Atm−/− p53−/− meiocytes. (a) Leptotene spermatocyte with faintly stained fragments of axial elements. (b) Zygotene nucleus with long axial elements and stronger SCP3 signal spots, which indicate some SC formation. (c) Aberrant spermatocyte I with fragmented SCs. Absence of H1t fluorescence indicates that this spermatocyte has not reached mid-pachytene. (d) Late-pachytene spermatocyte (arrowhead) with H1t-positive chromatin (red). (e) Diplotene spermatocyte with faintly labeled axial cores embedded in H1t-positive chromatin. (f) Nucleus of a round spermatid with a single DAPI-bright chromocenter and H1t-positive chromatin. DNA was stained with DAPI.

Harry Scherthan, et al. Mol Cell Biol. 2000 October;20(20):7773-7783.
3.
FIG. 2

FIG. 2. From: Meiotic Telomere Distribution and Sertoli Cell Nuclear Architecture Are Altered in Atm- and Atm-p53-Deficient Mice.

Histone H1t (red) and anti-SCP3 (green) immunostaining to Atm−/− testis nuclei. (a) An aberrant spermatocyte I shows axial cores and fragments of SCs near a large chromocenter (bright blue). (b) Spermatocyte nucleus at zygotene equivalent stage with fragments of SCs is devoid of H1t signals. The SEC nucleus below (arrowhead) displays a large nucleolus (dark area void of H1t signal) and distinct dispersed H1t signals throughout its chromatin. Note that H1t signals were not seen in wild-type SECs (not shown). (c) A rare late-pachytene spermatocyte nucleus exhibiting H1t fluorescence and strongly labeled SCs with thickened ends. DNA is counterstained with DAPI (blue).

Harry Scherthan, et al. Mol Cell Biol. 2000 October;20(20):7773-7783.
4.
FIG. 4

FIG. 4. From: Meiotic Telomere Distribution and Sertoli Cell Nuclear Architecture Are Altered in Atm- and Atm-p53-Deficient Mice.

Combined telo-FISH (FITC, green) and H1t immunostaining (Cy3, red) to suspension nuclei of Atm−/− p53−/− testis. (a) Premeiotic nucleus, which exhibits telomere signals (green) dispersed throughout the nuclear lumen, as seen at the focal plane at the nuclear center. (b) Top view of the nucleus of a bouquet spermatocyte discloses clustered telomeres at a limited sector of the nuclear periphery. This spermatocyte is most likely at leptotene-zygotene-equivalent stage, since H1t signals are absent. (c) A more advanced spermatocyte which displays faint H1t signals in its chromatin and a relaxed but still locally restricted accumulation of peripheral telomeres. Focal plane is at the top of nucleus. (d) H1t-positive spermatocyte nucleus (late pachytene or diplotene) exhibits dispersed telomeres. Focal plane is at top of nucleus. (e) The same nucleus as in panel d, but focal plane at the maximum nuclear diameter is shown. Telomeres are distributed over the nuclear periphery. (f) Two spermatid nuclei encountered in the Atm−/− p53−/− mouse testis show H1t fluorescence in their chromatin and formation of chromocenters. Nuclear DNA is counterstained with DAPI (blue).

Harry Scherthan, et al. Mol Cell Biol. 2000 October;20(20):7773-7783.
5.
FIG. 5

FIG. 5. From: Meiotic Telomere Distribution and Sertoli Cell Nuclear Architecture Are Altered in Atm- and Atm-p53-Deficient Mice.

IF staining of ATM (FITC, green) and SCP3 (red) in testis suspension cells of wild-type (a to c) and Atm−/− mice. Strong granular ATM IF signals are dispersed throughout the nuclei of (a) spermatocytes, (b) spermatids, and (c) SECs. Elongated sperm nuclei (c, right detail) do not exhibit ATM epitopes. ATM signals are reduced in the heterochromatin clusters of an SEC nucleus (c, dark-staining regions), while ATM is present in the heterochromatin clusters of a round spermatid (b). (d) ATM (green) and histone H1t (red) co-IF to wild-type spermatid nuclei. The round spermatid to the right shows extensive colocalization of ATM and H1t signals (yellow in the merge). The elongated spermatid nucleus in the center exhibits H1t but no ATM signals, while the condensed nucleus of a sperm head is void of IF signals. Atm−/−, an Atm−/− spermatocyte (nucleus to the left, identified by SCP3-positive SC fragments [red]) and an SEC (as identified by vimentin [Vim.] staining around its nucleus; right detail in red channel) lack ATM immunofluorescence. The autofluorescence in the green channel seen at the SEC position results from weak bleedthrough of the strong vimentin signal upon prolonged exposure. DAPI was used as the DNA counterstain.

Harry Scherthan, et al. Mol Cell Biol. 2000 October;20(20):7773-7783.
6.
FIG. 6

FIG. 6. From: Meiotic Telomere Distribution and Sertoli Cell Nuclear Architecture Are Altered in Atm- and Atm-p53-Deficient Mice.

SECs from control (a and b) and Atm−/− p53−/− mouse testis (c and d) stained for vimentin (TRITC; red) and hybridized with a FITC-labeled telomere PNA probe (fluorescein, green). (a to d) SECs positive for vimentin intermediate filament marker. (a) SEC with two chromocenters (blue) which are associated with one or two distinct telomere signals. (b) Vimentin-positive SEC with three chromocenters, each associated with one telomere signal. (c) Two-chromocenter Atm mutant SEC which shows two telomere signals at the larger chromocenter. (d) Mutant SEC with numerous heterochromatin clusters and telomere signals. The bar in d represents 10 μm and applies to a through d. (e) Wild-type two-chromocenter type I SEC with numerous minor-satellite (red) and associated telo-FISH signals (green). The inset shows an enlarged mouse metaphase chromosome (blue) with FISH signals of the minor-satellite probe (red) at the kinetochore region. (f) Nucleus of a wild-type two-chromocenter type II SEC which exhibits one strong minor-satellite signal (red) and an associated single-telomere signal (yellow) at each DAPI-bright chromocenter. DNA was counterstained with DAPI (blue). The bar in f represents 10 μm and applies to e and f.

Harry Scherthan, et al. Mol Cell Biol. 2000 October;20(20):7773-7783.

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