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. Author manuscript; available in PMC: 2024 Sep 5.
Published in final edited form as: J Bone Miner Res. 2019 Dec 30;35(4):698–713. doi: 10.1002/jbmr.3929

Figure 1.

Figure 1.

Screening for osteoblast inducing factors. (A) Schematic representation of the strategy to test osteoblast inducing factors. (B) 8 days after transduction of lentivirus containing six candidate transcription factors, live cell GFP image demonstrates Col2.3GFP expression. Scale bars: 1000 μm. Lentivirus containing pLV-tet empty vector was used as mock control. (C) 25 days after transduction of lentivirus containing various combination transcription factors, representative Alizarin Red S and Von kossa staining images in reprogrammed cells. (D) 25 days after transduction, flow cytometry analysis Col2.3GFP+ percentage in reprogrammed cells. All data represent mean ± SD (n=6; One-way ANOVA; *, P=0.006; +, P=0.005; ^, P=0.026; #, P<0.001; relative to mock). (E) 25 days after transduction, qRT-PCR analysis with osteogenic specific markers in reprogrammed cells. All data represent mean ± SD (For Col1a1 expression: n=5 for mock, Runx2, Runx2+Dlx5, or Runx2+Dlx5+Osx+ATF4 group; n=4 for Runx2+Dlx5+Osx group; n=3 for Runx2+ATF5, Runx2+Osx, or Runx2+Osx+ATF5, Runx2+Dlx5+ATF4 group; For Bglap expression: n=3; For Ibsp expression: n=5 for mock, Runx2, Runx2+Dlx5, Runx2+Dlx5+Osx, or Runx2+Dlx5+Osx+ATF4 group; n=3 for Runx2+ATF5, Runx2+Osx, or Runx2+Osx+ATF5, Runx2+Dlx5+ATF4 group; One-way ANOVA; **, P=0.006; +, P=0.012; *, P=0.046; #, P<0.001; ^, P=0.011; relative to mock). See also Figure S1–S4.