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. 2017 Oct 23;292(50):20657–20668. doi: 10.1074/jbc.M117.793885

Figure 4.

Figure 4.

1,25D directly regulates CD274 and CD273 gene expression via both VDREs. A–D, ChIP analysis of the effects of 1,25D on recruitment of the large subunit of RNA polymerase II (Pol 2) to the VDREs (A and B) and TSSs (C and D) in SCC25 cells and THP-1 cells, as indicated. E and F, effects of 1,25D on recruitment of the VDR to the TSS of CD274 or CD273 genes in SCC25 (E) and THP-1 cells (F), as indicated. The -fold change is calculated relative to the nonspecific IgG IP performed with the control sample. G, 1,25D treatment for the indicated times stimulates eRNA synthesis upstream of VDRECD273 + 47,959. Directed RT-qPCR was employed to show 1,25D-dependent production of eRNA 5′ of VDRECD273 + 47,959 and centered at 47,810 bp downstream of the CD273 TSS in THP-1 cells. C, vehicle (24 h); D, 100 nm 1,25D (24 h). A–F, n = 3; G, n = 5; ns > 0.05 ≥ * ≥ 0.01 ≥ ** ≥ 0.001 ≥ ***. Error bars, S.D.