Figure 5

A-D Flow cytometry sorting of peripheral blood mononuclear cells (PBMC) subpopulations: CD3/CD19, CD14, and CD16. PBMC were incubated with an antibody cocktail of CD3-Cy5, CD19-APC, CD16-FITC, and CD14-PE, then placed in the cell sorter. This method showed 93.7%, 96.4% and 96.5% purity for CD16, CD14 and CD3/19 fractions respectively. E-H. CK19 and HER2 signal was detected in sorted subpopulations of Peripheral Blood Mononuclear Cells. RNA from subpopulations of flow cytometry sorted cells was isolated and subjected to quantitative RT-PCR for CK19, HER2, and B2M. HER2 and CK19 expression/cell was calculated as the ratio of HER2/CK19 cell equivalents over the total number of PBL cells within the sample (determined using B2M expression). E. CK19 expression in sorted normal PBL. F. CK19 expression/cell in sorted normal PBL. G. HER2 expression in sorted normal PBL. H. HER2 expression/cell in sorted normal PBL