Expression of CD59 and CD105 on endothelial cells and apoptotic bodies in HUVEC culture treated with camptothecin Confluent HUVEC at 48 hrs in culture were treated for 24 hrs with 0.5% DMSO, 500 nM or 5 μM CPT in medium. After treatment the cell culture supernatant was pooled with harvested adherent cells. Mixture of cells, apoptotic bodies and other cell fragments was labeled with propidium iodide (PI) or with PE-labeled Mabs to CD59 or CD105 and analyzed by flow cytometry. Forward scatter (FSC) vs. side scatter (SSC) plot (A) shows gated intact single cells (G1) and apoptotic bodies and other cell fragments (G2) in a sample treated with 0.5% DMSO alone. PI – fluorescence vs. FSC plots of this sample is shown (B). Histograms showing expression of CD105 and CD59 on cells (C) and cell fragments (D) from HUVEC treated with 0.5% DMSO (solid line), 500 nM CPT (dotted line) or 5 μM CPT (dashed line) are shown. Binding of the IgG2a isotype control (IgG-IC) corresponded to the fluorescence of nonlabeled cells and cell fragments. Results are representative of three experiments.
Šimák et al. BMC Cell Biology 2002 3:11 doi:10.1186/1471-2121-3-11