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. 2019 Mar 18;1(1):56–66. doi: 10.1089/bioe.2018.0005

FIG. 6.

FIG. 6.

STC1 gene expression is upregulated with depolarization, and STC1 knockdown rescues early OS differentiation markers, but not calcification, in depolarized hMSCs. STC1 expression (A) and sodium phosphate cotransporter SLC20A1 expression (B) were quantified on day 7 by qPCR in OS-differentiated cells cultured under depolarizing (HK) or nondepolarizing (OS) conditions with or without 0.5 mM Pi supplementation. (C) STC1 expression was quantified 24 h after STC1 knockdown by siRNA. STC1-knockdown cells and negative control knockdown cells were then cultured in OS medium, OS medium with high K+ (HK), or control medium (CON). On day 7, cultures were evaluated for expression of STC1 (D), ALPL (E), or IBSP (F). Data points are mean normalized expression ± standard deviation, n = 4–6. (G, H) On day 21, cultures were evaluated for total calcium content. Data points represent mean normalized calcium content (μg Ca/μg DNA) ± standard deviation, n = 4. Different letters over bar graphs represent statistically different groups as determined by one-way ANOVA and the Tukey–Kramer post hoc test, p < 0.05. Asterisks (*) represent statistical differences between STC1-knockdown versus negative control knockdown cells, as determined by the Student's t-test, p < 0.05. ALPL, alkaline phosphatase gene; STC1, Stanniocalcin 1.