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. Author manuscript; available in PMC: 2020 Dec 1.
Published in final edited form as: Mol Cancer Ther. 2020 Mar 27;19(6):1266–1278. doi: 10.1158/1535-7163.MCT-19-0174

Figure 3. BMP4 enhanced OSB differentiation.

Figure 3.

A) OSB were co-cultured with Bo-786 cells in 3D hydrogels and were treated with or without recombinant BMP4 (100 ng/mL). BMP4 was added with each change of medium every other day. ALP mRNA and OSC mRNA were determined by real-time RT-PCR. ALP protein levels were determined by ALP activity assay. OSC protein levels were determined using OSC ELISA kit. Calcium deposits were determined by calcium assay. *: P < 0.05; **: P < 0.01 as compared to its vehicle control. B) BMP4 mRNA in BMP4-overexpressing Bo-786 cells (Bo-786/BMP4) were determined by real-time RT-PCR. The vector-expressing Bo-786 cells (Bo-786/Vector) were used as control. Secreted BMP4 protein levels in cell culture medium were determined using BMP4 ELISA kit. C) OSB were cultured in 2D system in ODM and treated with or without conditioned medium from Bo-786/BMP4 or its control Bo-786/vector cells for 14 days followed by ALP staining and ALP activity assay. D) OSB were co-cultured with Bo-786/BMP4 cells or with control Bo-786/Vector cells. ALP and OSC mRNA and protein expression and calcium deposits were determined as in (A). *: P < 0.05; **: P < 0.01 as compared to its vector control.