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. 2015 Feb 18;10(2):e112716. doi: 10.1371/journal.pone.0112716

Figure 5. Effects of S252W mutation on the osteogenic differentiation and mineralization of BMSCs.

Figure 5

(A) ALP staining showed less crystal violet-staining cells in cultured Fgfr2 S252W/+ BMSCs than in wild-type BMSCs on days 4, 7, and 14. (B) There was significantly less ALP activity (normalized to the total protein content of the sample, 562 nm) in Fgfr2 S252W/+ BMSCs. (C) Alizarin red staining of the mineralized osteoblasts showed more mineralized nodules on day 21 after osteogenic differentiation in Fgfr2 S252W/+ mice. (D) Bound alizarin red was dissolved with 5% SDS, 0.5 N HCl and measured at 415 nm to quantify the mineral content. Cultured Fgfr2 S252W/+ BMSCs showed more mineral content. (E–H) Relative expression levels of osteogenic marker genes were measured using qRT–PCR. The levels of oc, op, Runx2 and osteorix mRNA expression in differentiated BMSCs were markedly lower in Fgfr2 S252W/+ BMSCs on days 4, 7, and 14, but markedly higher on day 21. The expression levels of Col1 mRNA in differentiated BMSCs were markedly lower in Fgfr2 S252W/+ BMSCs on day 4, but they were higher on day 7 and markedly higher on days 14 and 21. Graphs show mean value ±SD (Student's t-test, *P<0.05, **P<0.01, ***P<0.001).