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. 2019 Jul 25;28(15):1004–1014. doi: 10.1089/scd.2019.0124

FIG. 4.

FIG. 4.

Periodontal AB fibroblasts upregulate terminal mineralization markers upon SFRP1 inhibition independent of RUNX2 and SP7. In this study, AB fibroblasts were subjected to mineralizing conditions with either induction media or in the presence of the SFRP1 inhibitor WAY-316606 at various concentrations. ALP activity was assessed after 7 days (A, top panel), while mineral deposits were detected using ARS after 14 days of culture (A, bottom panel). (B) Western blot analysis for expression of mineralization related genes in AB progenitors grown under osteogenic conditions with or without 1 μM WAY-316606 for 7 days. β-actin was used as a loading control. (C) Relative mRNA expression of osteogenic differentiation markers in AB fibroblasts grown in mineralization media with or without 1 μM WAY-316606 for 7 days. (D) mRNA expression of mineralization-related marker genes in AB cells stably expressed in con shRNA or SFRP1 sh1 shRNA. Expression was normalized against GAPDH, and statistical significance was determined using Student's t-test. (*P < 0.05, **P < 0.01). ARS, alizarin Red S; ALP, alkaline phosphatase; con, control; ind, induced.