Inhibition of VSMC proliferation in coculture requires uPA/uPAR signaling. VSMCs on coculture with monocytes were treated with BrdU. Incorporation was detected by immunofluorescence using monoclonal antibody against BrdU. Cells were counted using Hoechst 33258 nuclear staining. Representative results of n experiments are shown. (A) Result (n = 4) using wild-type VSMCs and wild-type monocytes. (B) uPA^-/- monocytes and wild-type VSMCs were used for coculture (n = 3). (C) uPAR^-/- VSMCs and wild-type monocytes were used (n = 3). (D) uPA^-/- VSMCs and wild-type monocytes were used (n = 3). The coverslips were counted in one given field for the number of cells in blue filter for Hoechst 33258 and for number of proliferating cells in red filter for BrdU (Alexa 546). Cells were counted (mean ± SD of 5-7 fields).