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Open Access Highly Accessed Research article

ARTEMIS stabilizes the genome and modulates proliferative responses in multipotent mesenchymal cells

Sarah A Maas12, Nina M Donghia1, Kathleen Tompkins13, Oded Foreman4 and Kevin D Mills15*

Author Affiliations

1 The Jackson Laboratory, 600 Main Street, Bar Harbor ME 04609, USA

2 Adnexus Therapeutics, Waltham, MA 02453, USA

3 Division of Infectious Diseases, University of North Carolina, Chapel Hill, NC27599, USA

4 The Jackson Laboratory, 4910 Raley Road, Sacramento CA, 95838, USA

5 Main Medical Center Research Institute, Scarborough Maine, USA

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BMC Biology 2010, 8:132  doi:10.1186/1741-7007-8-132

Published: 27 October 2010

Additional files

Additional file 1:

Aneuploidy without translocations in Art-null sarcomas. Spectral karyotype (SKY) analysis of Art-null sarcomas: (a) AP812, osteosarcoma; and (b) APJ4631, rhabdomyosarcoma. Shown for each is the 4',6'-diamidino-2-phenylindole (DAPI)-stained metaphase (inverted image, top left) with superimposed chromosome contours (blue), spectral image of SKY painted metaphase spread (top, middle), and computer classified image (top, right), as well as the karyotype table showing aneuploidy (bottom).

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Additional file 2:

The Art/Dclre1c, encoding ARTEMIS, is transcribed in mesenchymal stem cells (MSCs). (a) Schematic of reverse transcriptase polymerase chain reaction (RT-PCR) strategy to detect Art transcript in wild-type (WT) versus ArtΔ/Δ MSCs. PCR product detecting exons 1-4 (Art ex1-4) is common to both the WT and ArtΔ/Δ alleles (because the knockout allele eliminates exons 5-6. PCR product detecting exons 1-5 (Art ex1-5) is only amplified from WT cells, but not ArtΔ/Δ cells. (b) RT-PCR reactions detecting Art ex 1-4, Art ex 1-5, or glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (control) transcripts as indicated. Shown are data for either WT or ArtΔ/Δ fibroblasts or MSCs (as indicated beneath). These data confirm detection of Art ex 1-4 in both WT and ArtΔ/Δ MSCs, but detection of Art 1-5 only in WT MSCs. This confirms transcriptional expression of Art in MSCs and verifies the expected knockout in MSCs from ArtΔ/Δ mice.

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Additional file 3:

Control for differentiation specificity of WT or ArtΔ/Δ MSCs. (a) Fixed ArtΔ/Δ and Art MSCs treated with adipocyte- and osteocyte-specific differentiation medium were stained with the fluorescent lipid binding dye LipidTOX. Cells grown in osteocyte-specific medium are not positive for LipidTOX staining, indicating the absence of adipocytes in these culture conditions. (b) Fixed ArtΔ/Δ and Art MSCs treated with adipocyte- and osteocyte-specific differentiation medium were stained with the mineralized bone-specific stain alizarin red. Cells treated with adipogenic medium do not stain with alizarin red, indicating that mineralized bone is not present in adipogenic-treated cells.

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Additional file 4:

Photomicrograph of WT MSC culture following 2 days of serum withdrawal.

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Additional file 5:

Photomicrograph of WT MSC culture following 6 days of serum withdrawal.

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Additional file 6:

Photomicrograph of Art-null MSC culture following 2 days of serum withdrawal.

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Additional file 7:

Photomicrograph of Art-null MSC culture following 6 days of serum withdrawal.

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Open Data