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. Author manuscript; available in PMC: 2015 Mar 28.
Published in final edited form as: Circ Res. 2014 Feb 13;114(7):1094–1102. doi: 10.1161/CIRCRESAHA.114.302968

Figure 2. Inhibition of OGA increases O-GlcNAcylation and calcification in VSMC.

Figure 2

A) and B) Thiamet-G increased O-GlcNAcylation in VSMC. VSMC were treated with Thiamet-G for 6 hours at 0-10 μM, and Western blot was performed to determine O-GlcNAcylation. Representative blots from 4 independent experiments are shown in A. B) The intensity of the bands in each condition in A was quantified by NIH ImageJ and compared with that in the control (Thiamet-G, 0), defined as 1. C) and D) Thiamet-G induced VSMC calcification. VSMC were treated with osteogenic medium alone (Control) or osteogenic medium with 10 μM Thiamet-G for 3 weeks. Calcification was determined by Alizarin red staining (C) or quantified by Arsenazo III assay (D, n=3, *p<0.001). E) and F) Thiamet-G increased the expression of Runx2 and osteogenic marker genes, as determined by Western blot (E) and Real-time PCR analysis (F) in parallel experiments as in C and D (n=3, *p<0.01 compared to control).