Supplementary Information A mouse model of the ATR-Seckel Syndrome reveals that replicative stress during embryogenesis limits mammalian lifespan. Matilde Murga, Samuel Bunting, Maria F. Montaña, Rebeca Soria, Francisca Mulero, Marta Cañamero, Youngsoo Lee, Peter J. McKinnon, Andre Nussenzweig and Oscar Fernandez- Capetillo. Nature Genetics: doi:10.1038/ng.420
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Supplementary Information - Nature Research · Figure S4. Impaired oogenesis in ATRS/S mice. a, Immunohistochemistry of ATR+/+ and ATRS/S ovaries with an oocyte recognizing antibody
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Supplementary Information
A mouse model of the ATR-Seckel Syndrome reveals that replicative stress
during embryogenesis limits mammalian lifespan.
Matilde Murga, Samuel Bunting, Maria F. Montaña, Rebeca Soria, Francisca Mulero, Marta
Cañamero, Youngsoo Lee, Peter J. McKinnon, Andre Nussenzweig and Oscar Fernandez-
Capetillo.
Nature Genetics: doi:10.1038/ng.420
Nature Genetics: doi:10.1038/ng.420
Figure S1. Cranial anomalies in Seckel mice. a, Side and top views of CT scans
of 3 month old ATR+/+ and ATRS/S littermates. The skull surface of the mutant
animals shows less marked osseous relieves, with the indentation of the sutures
between the cranial bones being significantly reduced. In addition, the mutant
animals presented dental malocclusion which was due to a smaller curvature-
radius of the upper incisors (red arrowhead, 1), together with a rounding of the
crown of the lower incisors (red arrowhead, 2). The previous phenotypes were
100% penetrant. A deficient closure of the fontanelles was also frequently
detected (6/9 mice analysed). Scale bar indicates 2 cm. b, Head circumference
measurements in 3 month old ATR+/+ and ATRS/S mice. Note: A perimeter is
drawn around the head circumference of the animals for this analyses (see
dashed red lines in a). Due to the differences between the mice and humans, we
used the region of the skull that surrounds the brain, which is equivalent to the
occipitofrontal measurement done in humans. On average, Seckel animals
present a decrease on the head circumference of -8.81 SD, which is bigger to
that of height (-4.88 SD) or weight (-3.4 SD).
Nature Genetics: doi:10.1038/ng.420
Nature Genetics: doi:10.1038/ng.420
Figure S2. Neurological defects on ATRS/S mice. a, Picture of the brains from a
pair of one month old wt and Seckel animals. Scale bar indicates 0.5 cm. b, The
figure illustrates the generalized loss of cellularity (Niss1 staining, blue) and
acute depletion of astrocytes (GFAP staining, green) in the corpus callosum of 6
week old ATRS/S mice.
Nature Genetics: doi:10.1038/ng.420
Nature Genetics: doi:10.1038/ng.420
Figure S3. Analysis of the organ sizes of ATRS/S mice. a, Comparison of the
sizes of the different organs extracted from 3 month old ATR+/+ and ATRS/S
animals. The decrease in size of organs from the mutant animals is also
expressed in terms of the number of standard deviations below the wild type
average (last column; <>SD). b, ATR western blot in several organs from 1
month-old ATR+/+ and ATRS/S mice. ATR levels in newborns (p5) are also
shown for lung and testes.
Nature Genetics: doi:10.1038/ng.420
Nature Genetics: doi:10.1038/ng.420
Figure S4. Impaired oogenesis in ATRS/S mice. a, Immunohistochemistry of
ATR+/+ and ATRS/S ovaries with an oocyte recognizing antibody (DDX4). The
arrowhead points to the only maturing oocyte recognized in this section. Scale
bar (black) indicates 0.5 mm. b, Quantification of the number of DDX4 positive
cells found in a region of in ovaries from newborn and 1 month old animals. c,
Primary follicles of the newborn ATRS/S ovaries. Red arrows indicate the
presence of vacuoles. Scale bar (black) indicates 15 μm.
Nature Genetics: doi:10.1038/ng.420
Nature Genetics: doi:10.1038/ng.420
Figure S5. Functional complementation of ATR with the humanized murine
allele. a, PCR genotyping; and b, RT-PCR of ATR with primers at E8 and E10 in
littermate MEF lines. c, ATR western blot in several organs from ATR+/+ and
ATRHS/HS mice. d, A closer picture of the heads from a couple of 3 month-old
ATR+/+ and ATRHS/HS littermates. e, Weight of ATRHS/HS and control mice at 3
months of age (n=10).
Nature Genetics: doi:10.1038/ng.420
Nature Genetics: doi:10.1038/ng.420
Figure S6. Analysis of the frequency and functional capacity of Seckel HSCs.
a, Representative FACS plots from ATR+/+, ATRS/S and 18 month old ATR+/+