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. 2017 Sep 26;5:17044. doi: 10.1038/boneres.2017.44

Figure 7.

Figure 7

AFF4 is required for ID1 transcription and BMP2-induced responses. (a) Real-time RT-PCR shows altered expression of ID1. n=3. ***P<0.001. (b) Western blot analysis of ID1. (c) A ChIP assay for AFF4 shows that it binds to the promoter region of ID1. n=4. **P<0.01. (d) Relative luciferase activity after BMP2 (100 ng·mL−1) treatment for 6 h. Depletion of AFF4 blunts the BMP2-induced luciferase activity. n=4. ***P<0.001. (e) Real-time RT-PCR indicates that knockdown of AFF4 decreased SP7 expression after BMP2 (100 ng·mL−1) treatment. n=3. *P<0.05 and ***P<0.001. (f) Representative images of ALP staining. (g) Quantitative analyses of ALP activity. Knockdown of AFF4 decreases the ALP activity induced by BMP2 treatment. n=5. *P<0.05.