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Fig. 6. RNAi-induced loss of BMPR1A, by reducing MMP9 and MMP2 expression, attenuates proliferation of human vascular smooth muscle cells (HPASMCs). (A) Gelatin zymography using conditioned media of HPASMCs. Gelatin zymograms performed on conditioned media (left) and densitometric analysis (right) show MMP9 and MMP2 activities in SiBMPR1A-treated (SiBR1A) versus SiControl-treated (SiC) cells in response to serum (10% FBS) for 6 hours. Bars indicate mean±s.e.m. of densitometric values of SiBR1A proMMP9, MMP9, proMMP2 and MMP2 normalized to their corresponding SiC values (n=3-4). ^*P<0.05 and ^**P<0.01 between SiBR1A and SiC. (B) Proliferation of HPASMCs in response to serum using the MTT assay. HPASMCs transfected with SiC (white bars), SiBR1A, SiMMP9, SiMMP2 and combined SiMMP9 and SiMMP2 were subjected to 0.1% FBS (all white bars) or stimulated with 10% FBS (all black bars) for 72 hours, after which proliferation was assessed by the MTT assay. Bars indicate mean±s.e.m. of arbitrary OD₅₇₀ values normalized to values of SiC under 0.1% FBS. n=12 for siMMP2 and/or siMMP9 and n=26 for SiC and SiBMPR1A from three independent experiments. P<0.001 for serum-stimulated versus unstimulated comparisons, and ^***P<0.001 for comparisons with SiC at 10% FBS.

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