FIGURE 4.

Regulation of acetyl-CoA levels by AceCS1. A, WT MEFs were transfected with siRNAs against ACLY, AceCS1, or a nontargeting control. Cells were harvested 3 days after transfection for acetyl-CoA measurement, **, p < 0.01, n = 3. Error bars, S.E. B and C, control and AceCS1-knockdown mammary epithelial carcinoma cell lines were used for acetyl-CoA measurement (B); **, p < 0.01, n = 8; AceCS1 gene expression by quantitative PCR (C); ***, p < 0.00001, n = 9. D, MEFs were infected with lentiviruses expressing shRNA against a nontargeting control or against AceCS1. Several clones were screened, and clone 4 (showing maximum down-regulation of AceCS1 protein levels) was selected for further experiments. E, control (WT) and AceCS1-knockdown MEFs were synchronized by dexamethasone, cells were harvested at indicated time points, and gene expression was analyzed by real-time quantitative PCR. Gene expression was normalized to actin mRNA levels.