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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: J Thromb Haemost. 2014 Jul 17;12(8):1283–1293. doi: 10.1111/jth.12633

Fig. 1. 2bF8 transgene analysis.

Fig. 1

(A) Diagram of the pWPT-2bF8/MGMT construct. FVIII is driven by the platelet-specific αIIb promoter. MGMTp140k was driven by the MSCV promoter. (B) PCR detection of the 2bF8 transgene. DNA was purified from peripheral white blood cells. A 0.65 kb fragment from the 2bF8 expression cassette was amplified. Wild type (WT) mFVIII and mFVIIInull PCRs were used as controls to confirm that 2bF8/MGMT-transduced recipients were on the FVIIInull background. (C) qPCR determination of the average copy number of the 2bF8/MGMT transgene per cell in transduced recipients. Peripheral blood cell-derived DNA was analyzed for the 2bF8 transgene and normalized to the ApoB gene. These results demonstrate that 2bF8/MGMT genetically modified hematopoietic cells were viable and selectable by BG/BCNU treatments.