Fig. 3

Regulation of canonical Wnt signaling pathway by PKC/Akt axis. a–d DFCs were treated with 10 μM Akt activator SC-79 (a, c) or 200 nM Akt inhibitor MK2206 (b, d) for 15, 30, and 60 min. Expression of P-Akt (Ser473) (a, b) as control and P-GSK3β (Ser9) (c, d) was determined by Western blot analysis. e, f DFCs were cultivated in control medium (DMEM), osteogenic differentiation medium (ODM), or BMP2 containing differentiation medium with and without simultaneous treatment with 100 nM classical PKC inhibitor Gö6976 for 3 days. Expression of active β-catenin was determined in Western blot analysis after enrichment of cytoplasmic (e) or nuclear (f) proteins. Bar charts show means + SD (n = 3). One-way ANOVA was performed to compare all groups including Tukey’s post hoc tests comparing different groups in the same medium pairwise (a–d). Student’s t test was performed to determine statistically significant differences between the control and treatment group for each medium (e, f). *p < 0.05, **p < 0.01, ***p < 0.001