Figure 3.

Jarid2 recruits Ezh2 to an artificial promoter. (A) Schematic of the luciferase reporter under the control of the TK promoter, with Gal4 DNA-binding sites upstream. (B) Luciferase activity was measured 24 h after doxycyclin induction of the stably integrated transgene encoding Jarid2 fused to the Gal4 DNA-binding domain (Gal4-Jarid2). The values were normalized to the amount of protein. Error bars represent SD of three independent experiments. (Inset) Gal4-Jarid2 expression was checked by Western blot. Actin was used as a loading control. (C) ChIP experiments were performed before (−Dox) and after (+Dox) Gal4-Jarid2 induction. Chromatin was immunoprecipitated with antibodies against the proteins indicated at the bottom. DNA enrichment was analyzed at the luciferase transgene. Results are presented as fold of enrichment over control. Error bars represent SD of three independent experiments. (D) ChIP experiments were performed with or without induction of the stably integrated transgene encoding Gal4-Ezh2. DNA enrichment was analyzed at the luciferase transgene. (E,F) Same as C, but DNA enrichments were analyzed at the endogenous Otx1 (E) and Irx4 (F) promoters. Insets show the levels of Otx1 and Irx4 mRNAs, normalized to those of GADPH.