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Fig. 1 | Stem Cell Research & Therapy

Fig. 1

From: Classical isoforms of protein kinase C (PKC) and Akt regulate the osteogenic differentiation of human dental follicle cells via both β-catenin and NF-κB

Fig. 1

Inhibitory role of classical PKCs during osteogenic differentiation of DFCs and Akt activity during differentiation and after inhibition of classical PKCs. a Protein expression of PKCα after 1, 7, 14, or 28 days cultivation in control medium (DMEM), osteogenic differentiation medium (ODM), or BMP2 containing differentiation medium determined by Western blot analysis. b DFCs were cultivated for 28 days in ODM and concurrently treated with 100 nM inhibitor of classical PKCs Gö6976 for either the whole 28 days or only during the first, second, third, or fourth week of differentiation. Cells cultivated in DMEM were used as control. 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 are shown below the relative quantification results. c Protein expression of P-Akt (Ser473) in DFCs as an indicator of Akt activity after 7, 14, or 28 days cultivation in control or differentiation media as above, determined by Western blot analysis. Expression of total Akt was determined as control. d, e Protein expression of P-Akt (Ser473) in DFCs after cultivation in control or differentiation media as above for 1 (d) or 7 (e) days and simultaneous treatment with 100 nM classical PKC inhibitor Gö6976, determined by Western blot analysis. Bar charts show means + SD (n = 3). One-way ANOVA was performed to compare different media at the same time point including Tukey’s post hoc tests comparing the individual differentiation media with the control medium (a, c) or to compare all groups including Tukey’s post hoc tests comparing differences between continuous Gö6976 treatment in ODM and other groups in ODM (b). Student’s t test was performed to determine statistically significant differences between the control and treatment group for each medium (d, e). *p < 0.05, **p < 0.01, ***p < 0.001

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