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Supplementary Methods Transgenic mice ApoE -/- mice (Taconic M&B, Ry, Denmark) and eGFP + mice (Jackson Laboratories, Bar Harbor, ME), both backcrossed more than ten times to C57BL/6 mice, were <strong>in</strong>tercrossed to obta<strong>in</strong> eGFP + apoE -/- mice (hemizygous for the eGFP transgene). 1,2 Genotyp<strong>in</strong>g for apoE was performed us<strong>in</strong>g primers recommended by the Jackson Laboratory (5-GCCTAGCCGAGGGAGAGCCG-3, 5- TGTGACTTGGGAGCTCTGCAGC-3, and 5-GCCGCCCCGACTGCATCT-3). Phenotyp<strong>in</strong>g for eGFP was done us<strong>in</strong>g a Woods UV lamp. Bone marrow transplantations Crude bone marrow was flushed from femurs and tibias from eGFP + apoE -/- mice 8 weeks of age (n=7). <strong>Cells</strong> were washed twice <strong>in</strong> RPMI-1640 (Invitrogen) and filtered through a 70 μm mesh. ApoE -/- mice (n=28) 8 weeks of age were irradiated with 10 Gy from a 137 Cs source and 10 7 unfractionated bone marrow cells were <strong>in</strong>jected <strong>in</strong>to a tail ve<strong>in</strong> 4 hours later. Mice were housed with filter tops and adm<strong>in</strong>istered oxytetracycl<strong>in</strong> (2g/l Terramyc<strong>in</strong> vet. 20%) one day prior to and 7 days after transplantation. Flow cytometry Blood (50 μl) was lysed <strong>in</strong> erythrocyte lysis buffer (NH4Cl 8.02 g/L, NaHCO3 0.84 g/l, Na2EDTA 0.37 g/l). The fraction of green fluorescent leukocytes <strong>in</strong> peripheral blood was measured <strong>in</strong> a forward-side scatter gate that was enriched for CD45 + cells (>98%) and excluded only a few CD45 + cells (~2%) as established <strong>in</strong> pilot experiments. Downloaded from http://atvb.ahajournals.org/ by guest on June 4, 2013
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