Figure 6.
CAU1 Regulates the Histone Methylation of the CAS Chromatin and the Consequent CAS Expression in Response to [Ca2+]o.
(A) A diagram of the genomic structure of CAS. Genomic regions A to E subjected to ChIP assay are indicated.
(B) and (C) ChIP assay with antibodies against H4R3sme2 (B) to determine histone methylation status or against GFP (C) to determine if CAU1 binds to the CAS promoter using 35S/EYFP:CAU1-cau1 plants. TUB8 was used as an internal control in (B). Three independent experiments were performed, and values are mean ± se.
(D) and (E) ChIP assay with antibodies against H4R3sme2 within the B region (D) or against GFP within the A region (E) of CAS. Wild-type and cau1 plants were treated with calcium as indicated. Three independent experiments were performed, and TUB8 was used as an internal control in (D). Values are mean ± se.
(F) Immunoblotting analysis of CAU1 levels in response to [Ca2+]o. Twenty-four-day-old plants were exposed to 0, 0.5, or 10 mM Ca2+ treatment for 4 d. Immunoblotting was performed as described in Methods. **P < 0.01.