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. 2024 Jan 4;8(2):ziad014. doi: 10.1093/jbmrpl/ziad014

Figure 2.

Figure 2

The effect of MerTK deficiency on osteoblast differentiation. (A) Calvarial cells isolated from WT or Mertk KO mice were stimulated with 30 ng/mL BMP2 for 7 d. Cells were then stained for alkaline phosphatase activity. Scale bar indicates 2 mm. (B) Calvarial cells were incubated with 30 ng/mL BMP2 for 14 d and subjected to alizarin red staining. Scale bar indicates 2 mm. (C) WT or Mertk KO calvarial cells were stimulated with 30 ng/mL BMP2 for 7 d. The mRNA expression levels of alkaline phosphatase (Alpl), Osterix (Sp7), and Osteocalcin (Bglap) were measured by real-time RT-PCR analysis. Data are mean of 4 experiments performed in triplicates. The P-values for two-way ANOVA in the order of interaction, genotype, and BMP2 stimulation were Alpl (<.001, <.001, <.001), Sp7 (<.001, <.001, <.001), and Bglap (.156, <.001, <.001). (D) Cell lysates in (C) were subjected to western blotting for the expression or phosphorylation of key regulators of osteoblast differentiation. (E) The band intensities in (D) were quantified using results from three independent experiments. The P-values for two-way ANOVA in the order of interaction, genotype, and BMP2 stimulation were β-catenin (.027, .002, <.001), pGSK (.991, <.001, .788), pSmad (<.001, <.001, .002), and NFATc1 (.099, <.001, <.001).