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. Author manuscript; available in PMC: 2019 Aug 1.
Published in final edited form as: Mol Cancer Res. 2018 May 14;16(8):1226–1240. doi: 10.1158/1541-7786.MCR-17-0406

Figure 2. Guadecitabine degrades DNMT1 in the absence of DNA synthesis and cell division.

Figure 2

(A). BrdU immunofluorescence labeling (2h) was performed in preadipocytes and adipocytes treated with the DNA synthesis inhibitor aphidicolin (APC) (1μg/mL, daily 4X). (B) MTT cell proliferation assay was performed in preadipocytes and adipocytes treated with aphidicolin (APC) (1μg/mL, daily 4X). (C). Propidium iodide cell cycle flow cytometry analysis was performed on preadipocytes and adipocytes treated with APC (1μg/mL, daily 4X). (D). Subcellular fractionation was performed on adipocytes treated with either guadecitabine (100nM, daily 3X) alone or APC (1μg/mL, daily 4X) starting on day 1 and guadecitabine together with APC therein after for 3 days. Western blot was performed on cytoplasmic and chromatin fractions and probed for DNMT1, histone H3, and GAPDH. All experiments were performed in triplicate. (*P < 0.05).