Figure 3. Expression of Notch1 and RBP-Jκ in VSMCs.
(A) mRNA level of Notch1 and RBP-Jκ were measured by real-time RT-PCR in VSMCs cultured in different media. (B) Protein level of Notch1 and RBP-Jκ were measured by Western blotting in VSMCs cultured in different media. Notch1 and RBP-Jκ significantly increased by 3 to 4 folds in VSMC cultured in osteogenic medium compared to control at both mRNA and protein levels, suggesting that the Notch1-RBP-Jκ signaling pathway is activated in osteogenic conversion of VSMCs. DAPT reduced the expression of Notch1 by 50% and RBP-Jκ by 30% in VSMCs compared to osteogenic cells at bothe mRNA and protein levels. Similarly, OPG reduced the expression of Notch1 and RBP-Jκ compared to VSMCs in the osteogenic medium in a dose dependent manner. *, P<0.05 vs control, #, P<0.05 vs osteogenic medium, and &, P<0.05 vs OPG 0.1ng/ml. n=3.