Fig. 2. Effect of PDGF and bFGF on UPA expression in human vascular SMC after 24 and 48 hours stimulation. Growth-arrested subconfluent SMC were cultured in DMEM/Ham's F-12 with 0.2% FCS or with 1% FCS in the absence or presence of PDGF (20 ng/ml) and bFGF (10 ng/ml). (A) UPA antigen levels in cell lysates. Data from a minimum of six independent experiments performed in triplicates are given as box-and-whisker plots; boxes denote interquartile ranges with median values shown as horizontal lines inside the boxes. Whiskers denote ranges; outliers beyond 1.5-times interquartile ranges are represented by individual symbols ([UNK]). UPA antigen levels in PDGF- or bFGF-stimulated cells were significantly higher than in cells treated with 1% FCS alone both after 24 hours (P=0.03 for PDGF and P=0.001 for bFGF; Mann-Whitney test) and 48 hours of stimulation (P<0.001 for PDGF and P<0.001 for bFGF; Mann-Whitney test). (B) UPA transcripts in unstimulated (1% FCS) and PDGF- or bFGF-stimulated SMC were analyzed by RT-PCR after 24 and 48 hours treatment. Levels of UPA messenger RNA were estimated by normalization against co-amplified glyceraldehyde-3-phosphate dehydrogenase (GAPDH); M, size marker.