G9a knockdown decreases metastasis-related signaling. A. Quantitative RT–PCR verification of the expression of genes identified in the microarray: control shRNA (blue) versus G9a shRNA1- and shRNA2-expressing cells (red and green, respectively). Upper panel: genes down-regulated after G9a depletion; Lower panel: genes up-regulated after G9a depletion. Genes analyzed shown on x-axis. B. OV-90 cells transfected with long-form or short-form G9a for 48 h and harvested for RNA extraction. G9a-regulated genes identified from microarray as indicated were examined by quantitative RT–PCR. C. G9a binding and H3K9 dimethylation regions of the promoters determined by ChIP assay in SKOV-3 cells. Schematic diagram of the human PPP1R15A promoter (GenBank accession number NC_000019) (Top) and CDH1 promoter (GenBank accession number NC_000016) (Bottom) shown. Numbered arrows indicate primer sets used for ChIP analysis. D. Akt and ERK phosphorylation status and E-/N-cadherin expression examined in control and G9a knock down cells. Results of quantitative RT–PCR are mean values of three independently repeated experiments done in duplicate and are displayed as mean ± SD. *P < 0.05, **P < 0.01, indicate a statistically significance by one way ANOVA followed by Bonferroni test.