Figure 4: Effect of methotrexate on serum-stimulated proliferation of human vascular smooth muscles cells assessed by cell counting. Cells were seeded at low density in 30 mm diameter plates and grown in DMEM with 10% fetal calf serum (FBS). After 3 days cells were transferred to 0.1% FBS medium or 5% FBS medium with or without Methotrexate (0.1–10 μM). Cells in all dishes were examined visually then counted after 3 days. The experiment was repeated three times with near identical results. Results are expressed as mean ± SEM of cell count from 4 separate dishes. Statistical analysis was performed using one-way ANOVA, ## P<0.01 (0.1 % FBS vs 5 % FBS) and ** P < 0.01 (5 % FBS vs Methotrexate).