Figure 8.

CyPA promotes migration and proliferation of mouse aortic smooth muscle cells (MASM). (A) Migration of MASM in response to 10% fetal bovine serum (FBS) in Boyden chamber assay. Tg-MASM, WT-MASM and KO-MASM were starved overnight and then seeded in the upper Boyden chamber on collagen-precoated PVP-free polycarbonate membranes. FBS (10%) was added to the lower chamber. Migration was determined and the maximum increase was normalized to 100%. (B) WT-MASM were starved overnight and then seeded in the upper Boyden chamber. Tg-CM, WT-CM or KO-CM was added to the lower chamber. Cells were incubated for 8 hours at 37 °C in a 95% air/5% CO2 humidified incubator. The membranes were removed, and cells were stained. The relative increases in cell number were determined by quantitative densitometry. ⋆P<0.01. Data are mean ± SD. n = 6 in each group. (C) Conditioned medium (CM) from VSMC-Tg (Tg-CM) or Control (Cont-CM) promotes cell proliferation. WT-MASM were seeded in 96-well plates in DMEM supplemented with 10% FBS, serum starved for 24 hours, and stimulated with Tg-CM or Cont-CM for 5 days. CM was changed at day 3 and cells were counted at day 2 and day 5. Data are mean ± SD. ^†P<0.05. (D-F), Effect of PDGF-BB and FBS on proliferation of Tg-MASM and Control-MASM. After starvation for 24 hours, MASM were incubated with DMEM (D) or stimulated with 25 ng/mL PDGF-BB (E) or 10% FBS (F) for 5 days. Medium was changed at day 3 and cells were counted at day 2 and day 5. Data are mean ± SD. ⋆P<0.01. n = 8 in each group.