FIGURE 1.

Selective positive and negative effects of G9a depletion on E₂-regulated gene expression. MCF-7 cells were transfected with SMARTpool siRNA targeting G9a (siG9a) or siRNA against a nonspecific sequence (siNS). Cells transfected with siRNA were cultured for 3 days in hormone-free medium prior to addition of 100 nm E₂ for the indicated time. A, whole cell extracts from cells at the zero time point of E₂ treatment were analyzed by immunoblot with antibodies for G9a and actin. B, total cellular RNA was extracted from cells at the zero time point of E₂ treatment and used to measure G9a mRNA level by qRT-PCR. C, total cellular RNA was extracted from cells at the indicated time point of E₂ treatment and analyzed by qRT-PCR to determine levels of the indicated mRNAs, which are normalized to the level of GAPDH mRNA. Results shown are the mean and S.D. for three PCR reactions performed with cDNA samples from a single experiment, and are representative of at least three independent biological experiments performed on different days.