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. 2017 Sep 25;18(10):2053. doi: 10.3390/ijms18102053

Figure 6.

Figure 6

APE1/Ref-1 overexpression inhibits inorganic phosphate (Pi)-induced alkaline phosphatase (ALP) activity and Runx-2 and Pit-1 expression in vascular smooth muscle cells (VSMCs). (A) ALP activity was determined. At the end of incubation, cells were washed with phosphate-buffered saline and solubilized with 1% Triton X-100 in 0.9% NaCl, and ALP activity was normalized to protein concentration. Cells were treated with Pi (5 mM) for 6 days, and VSMCs were infected with an adenovirus encoding APE1/Ref-1 at a multiplicity of infection (MOI) of 100, with Adβgal used at the same MOI as a control. Each bar represents the mean ± standard error of the mean (n = 5). ** p < 0.01 vs. control; # p < 0.05 vs. Pi alone by one-way ANOVA; (B) APE1/Ref-1 inhibited Pi-induced Runx2 and Pit-1 expression in VSMCs. Changes in Runx2 and Pit-1 expression in VSMCs were evaluated by Western blot analysis. VSMCs were infected with an adenovirus encoding APE/Ref-1 at an MOI of 300, with Adβgal used at the same MOI as a control. Similar results were observed in replicate experiments, and representative data is shown.