Figure 3. CD36 promotes VSMC proliferation in vitro.
A. Assessment by cell counting. Equal number of VSMCs were seeded in 6 cm plates, cultured in DMEM containing 10% FCS and counted daily for 4 days (n=5/group). * p<0.05 WT vs. Cd36-/−. B. MTT cell proliferation assay. 105 cells/well were seeded onto a 6 well plate. For the basal control, the cell culture media was changed to serum-free media 6 hrs after plating (to allow adhesion) and cultured overnight. MTT assay was performed the next day (= day 0 control). For other time points, the MTT assay was performed exactly 24, 48 and 72 hrs after plating cells. Data are expressed as fold change over Day 0, n=9/group. C. CD36 is necessary for PDGF induced VSMC proliferation. Cells were seeded into a 96-well plate in a density of 103 cells/well and cultured for onvernight. The cells were then synchronized by incubation in serum-free DMEM for 24 hours, and then stimulated with indicated conditions. Cell proliferation were assessed by MTT assay. D. Cell cycle analysis. 5 × 105 cells were seeded into a 10 cm plate and cultured for 24 hrs. Cell cycle was then analyzed by flow cytometry using propidium iodide staining. n=3/group, * p<0.01 WT vs. Cd36-/−.