Fig. 2

Mineralization of DFCs after treatment with Akt activators/inhibitors and interactions with PKC and BMP signaling. a–d DFCs were cultivated for 28 days in control medium (DMEM), osteogenic differentiation medium (ODM) or BMP2 containing differentiation medium, and concurrently treated with either 10 μM Akt activator SC-79 (a), 200 nM Akt inhibitor MK2206 (b), or 100 nM classical PKC inhibitor Gö 6976 alone or in combination with SC-79 (c, d). Mineralization of extracellular matrix was determined by Alizarin Red staining. Microscopic photographs (total width of each photograph corresponds to 1.24 mm) of stained cells and relative quantification results are shown. e DFCs were treated with 10 μM Akt activator SC-79 for 15, 30, and 60 min. Protein expression of P-SMAD 1/5 (Ser463/465) was determined by Western blot analysis. Bar charts show means + SD (n = 3). Student’s t test was performed to determine statistically significant differences between control and treatment group for each medium (a, b). One-way ANOVA was performed to compare all groups including Tukey’s post hoc tests comparing different groups in the same medium pairwise (c–e) or DMEM to ODM/BMP2 control groups (c, d). *p < 0.05, **p < 0.01, ***p < 0.001