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. Author manuscript; available in PMC: 2018 Feb 14.
Published in final edited form as: J Cell Physiol. 2015 Sep 18;231(5):1001–1014. doi: 10.1002/jcp.25188

Fig. 8. Runx2 binding to OC/BGLAP2 gene promoter during early G1 is independent of α-amanitin.

Fig. 8

MC3T3-E1 cells were synchronized by incubation for 16 h with nocodazole to generate a mitotic block. Mitotic cells were pre-treated with or without the transcriptional inhibitor α-amanitin for 2 h and harvested by gentle agitation and then released through mitotic into G1 the addition of fresh culture medium supplemented with (+) or without (−) α-amanitin, and cells were harvested after 0, 2, 4, and 6 h. Cells were crosslinked with 1% formaldehyde and sonicated chromatin fragments were immune-precipitated with a Runx2 polyclonal antibody. The enrichment levels of OC/BGLAP2 gene promoter sequences (−276/−5 pb) in the precipitated chromatin were determined by QPCR using specific primers. Student’s t test was performed to assess the significance of the observed differences. *P = 0.024; **P = 0.002.