Figure 6. C/EBPα is required for YY1, Med12, and RAD21 loading in “assisted” regions.
- Left panel: ChIP experiments of C/EBPα in cells expressing either shControl or shC/EBPα. The hormone‐regulated genes associated with the “Assisted” regions and tested by RT–PCR were AZIN1, SRCAP, ZBED3, PPP1R14D, and DUSP1 (n = 5; left panel). The inset shows the extent of C/EBPα depletion, using LSD1 as loading control. Right panel: ChIP experiments of YY1, Med12, and CTCF in cells expressing either shControl or shC/EBPα at T0. Precipitated DNA was analyzed by PCR for the presence of sequences corresponding to the assisted regions localized at −1.3 kb from the DUSP1 gene. Data are represented as mean ± SD from three experiments performed in duplicate.
- ChIP experiments of YY1 and Med12 in cells expressing either shControl or shC/EBPα before and after exposure to hormone. Precipitated DNA was analyzed by PCR for the presence of sequences corresponding to the “assisted” regions (n = 5) as shown in panel (A). Data are represented as mean ± SD from three experiments performed in duplicate.
- ChIP experiments of RAD21 in cells expressing either shControl or shC/EBPα before and after exposure to hormone. Precipitated DNA was analyzed by PCR for the presence of sequences corresponding to the “assisted” regions (n = 5) as shown in panel (A) (left panel) and DUSP1 (right panel). Data are represented as mean ± SD from three experiments performed in duplicate.
- ChIP‐seq experiments of PR in cells expressing either shControl or shC/EBPα after exposure to hormone. Reads profile for “assisted”, “constitutive”, “induced shared”, and “random” regions are shown.
- DNase I accessibility assays in cells expressing shControl and shC/EBPα and treated or not with hormone (see Materials and Methods). Accessibility was tested in 5 “assisted” regions as described in panel (A) and in 5 “induced shared” regions associated with ACOTT, WTAP, MUC21, CREBL2, and CAMKK2 genes. Each value corresponds to the mean ± SD of three experiments performed in duplicate. **P < 0.05 using Student's t‐test