Expression of PGK-neo in β6I mice.
(A) Flow cytometric analysis was used to quantitate cells expressing immunoreactive neomycin phosphotransferase. Bone marrow cells were first labeled with PE-conjugated monoclonal antibodies specific for cell surface antigen Ter-119, Gr-1, or CD3. Cells were then fixed in Cytofix/Cytoperm solution and incubated with rabbit anti-Neo polyclonal antibody and fluorescein-conjugated antirabbit secondary antibody. Neo expression is plotted on the x-axis, and surface markers are plotted on the y-axis. (B) Bone marrow cells from β6I/β6I mice were analyzed for the presence of immunoreactive neomycin phosphotransferase by indirect immunofluorescence microscopy. Cells were first stained with PE-conjugated antibody specific for murine erythroid lineage marker Ter-119, spun onto coverslips, fixed in methanol, and incubated with rabbit anti-Neo polyclonal antibody and fluorescein-conjugated antirabbit secondary antibody. Nuclear DNA was visualized with DAPI. A representative microscopic field is shown for Neo (Neo), Ter-119 (Ter-119), and DNA staining (DNA). A merged micrograph of the Neo and Ter-119 labeling patterns is also shown (Merge). Arrows indicate, Ter-119+/Neo+ cells; arrowheads, Ter-119-/Neo+ cells. Magnification × 200.
