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. 2017 Jan 5;12(1):e0169225. doi: 10.1371/journal.pone.0169225

Fig 3. Long-term dexamethasone treatment inhibits the induction of ANGPTL4 transcription, together with down-regulation of the GR.

Fig 3

(A) Protocol for Long-Term Dexamethasone Treatment (LTDT). For LTDT, HepG2 cells were initially cultured in DMEM medium supplemented with 10% DCC-treated FBS and 100 nM dexamethasone (Dex) for 14 days. Black arrows show sampling times. (B) Decrease in ANGPTL4 induction in LTDT cells. (C) Decreased enrichment of GR after Dex treatment of LTDT cells. ChIP-qPCR analysis was performed using an anti-GR antibody and an anti-rabbit IgG (control), followed by quantitative PCR using specific primers for each AG site and the control (NC). (D) CTCF enrichment in control and LTDT cells. ChIP-qPCR analysis was performed using an anti-CTCF antibody and anti-rabbit IgG (control), followed by quantitative PCR using specific primers for each AC site. (E) Expression of CTCF and GR after Dex treatment of control and LTDT cells. The amount of GR decreased in most LTDT cells. The relative level of GR normalized to that of β-tubulin is shown below. Uncropped image of western blot analysis is shown in S6 Fig. Asterisks indicate statistically significance between control and LTDT cells at each time point. **P < 0.01, ***P < 0.005.