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Blood, Vol. 113, Issue 21, 5094-5103, May 21, 2009
Long-term polyclonal and multilineage engraftment of methylguanine methyltransferase P140K gene-modified dog hematopoietic cells in primary and secondary recipients
Blood Beard et al.
113: 5094
Supplemental materials for: Beard et al
Files in this Data Supplement:
- Table S1. Dogs transplanted with MGMT-P140K gene-modified cells (PDF, 52.1 KB)
- Table S2. Closest proto-oncogene promoter to unique integration sites before chemotherapy (PDF, 111 KB)
- Table S3. Closest proto-oncogene promoter to unique integration sites after chemotherapy (PDF, 91.5 KB)
- Table S4. RIS isolated both before and after chemotherapy (PDF, 25.7 KB)
- Figure S1. Multilineage repopulation of gene-modified cells in secondary recipients (JPG, 177 KB)
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Representative flow cytometry plots of a secondary recipient (black dog) showing (A) forward and side scatter of total cells, stained for (B) DM5 (E) CD14 (H) CD3, sorted for (C) DM5+ granulocytes, (F) CD14+ monocytes, (I) CD3+ lymphocytes, and (D, G, and J) back-gated to a forward and side scatter plot.
- Figure S2. Durable in vivo selection of multiple clones (JPG, 65.8 KB)
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Long-term follow-up of (A) G187 gene marking in granulocytes (closed circle) and lymphocytes (open circle), (B) G250 gene marking in granulocytes (closed circle) and lymphocytes (open circle), and (C) G258 gene marking in granulocytes (closed circle) and lymphocytes (open circle) before and after in vivo selection of MGMTP140K-GFP gene-marked cells. Treatment with O6BG and BCNU denoted by black arrows and treatment with O6BG and temozolomide denoted by hashed arrow. (D) A representative LAM-PCR gel of the most recent time point in all dogs. Letters below the gel indicate the enzyme used for digestion of the genomic DNA to attach the linker cassette during LAM-PCR (M: Msp I, T: Tsp509 I, and B: BspH I).
- Figure S3. Chemoprotection in secondary recipient (JPG, 28 KB)
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Absolute neutrophil count (ANC) closed circles and platelet count (PLT) open circles plotted as days after treatment with O6BG and BCNU in a secondary recipient. Day of treatment with O6BG and BCNU is day 0.
- Figure S4. In vivo selection in secondary recipient (JPG, 21.8 KB)
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Gene-modified WBC in secondary recipient as a function of time. Chemotherapy (O6BG/BCNU) denoted by small black arrow.
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