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

Fig. 6.

Fig. 6

Comparison of marker gene expression in osteoblasts and VSMCs. (A) Expression of Runx2, Sp7, Alpl, Bglap2 and Spp1 was increased in calcifying VSMCs compared to control VSMCs but, in all cases, mRNA expression levels were lower in VSMCs than osteoblasts. (B) Osteoblasts express mRNA for all the VSMC marker genes studied. Only Cald1, Myh10 and Smtn were expressed at a higher level in VSMCs than osteoblasts. mRNA expression is presented as the fold change in VSMC expression relative to bone-forming osteoblasts. Values are mean ± SEM (n = 4 RNA sets), *** = p < 0.001, ** = p < 0.01 * = p < 0.05. (C) Protein expression of Sm22α was lower in calcifying VSMCs compared to control VSMCs at both day 7 and 14; calcification had a limited effect on Acta2 levels. Osteoblasts also displayed strong expression of these proteins with Sm22α expression being higher in osteoblasts than VSMCs. Runx2 was detected in osteoblasts, control and calcifying VSMCs with the highest levels being seen in osteoblasts. OPN protein was expressed at all stages of osteoblast differentiation but protein levels were highest in mature, mineralising osteoblasts. OPN protein was only evident in calcifying VSMCs at day 14. Blots shown are representative of experiments performed 3 times with different protein sets.