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Figure 3


Fig. 3. The NO-cGMP-PKG pathway transcriptionally regulates MMP-13 expression during MC3T3-E1 differentiation. MC3T3-E1 cells were induced to differentiate and analyzed at the time points indicated. (A) MMP-13 promoter activity in MC3T3-E1 transiently transfected with pMMP-13 WT, and incubated with 100 µM DEA-NO, or 200 µM of the iNOS inhibitor 1400W (n=3, mean ± s.d., *P<0.05 versus control). (B) Measurement of MMP-13 promoter activity in MC3T3-E1 cells transiently transfected with pMMP-13 WT, and incubated with 100 µM DEA-NO, 10 µM 8-Br-cGMP, 20 µM ODQ, and 100 µM DEA-NO plus 20 µM ODQ (n=3, mean ± s.d., *P<0.05 versus control, #P<0.05 versus DEA-NO). (C) Measurement of MMP-13 promoter activity in MC3T3-E1 cells transiently transfected with pMMP-13 WT and co-transfected with a dominant-positive construct of PKG1-{alpha} (black bars) (n=3, mean ± s.d., *P<0.05 versus control). MMP-13 mRNA was evaluated in these cells by RT-PCR analysis using GAPDH as a control (inset shows a representative experiment and a densitometric analysis of three independent experiments, mean ± s.d., *P<0.05 versus absence of PKG).





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