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. 2019 Jul 1;380(1):100–113. doi: 10.1016/j.yexcr.2019.04.020

Fig. 4.

Fig. 4

TNAP activity is 100-fold higher in bone forming osteoblasts than calcifying VSMCs. (A) Representative whole well scans and phase contrast microscopy images of osteoblasts and VSMCs stained to visualise TNAP expression. Bone-forming osteoblasts displayed the highest level of TNAP expression with only limited staining in calcifying VSMCs (highlighted by the arrow). No obvious TNAP staining was observed in control VSMCs. Scale bars: phase contrast microscopy images = 250 μm, tissue culture wells = 0.5 cm. (B) Rank order of TNAP activity was bone-forming osteoblasts >>> calcifying VSMCs > control VSMCs. TNAP activity was 50% higher in calcifying VSMCs compared to control VSMCs. (C) Rank order of total NPP activity was control VSMCs > calcifying VSMCs > bone-forming osteoblasts. (D) Culture with a TNAP inhibitor (≥0.1  μM) reduced bone formation by up to 90%. (E) Inhibition of TNAP had no effect on VSMC calcification. Values are mean ± SEM (n = 6 replicate wells), *** = p < 0.001.