Fig. 8.
Suppression of ASAH1 affects histone H3 acetylation levels. A, H295R WT and ASAH1KD (pretreated with 5 μg/ml tet for 96 h) cell lysates were isolated and separated by SDS-PAGE. Acetyl-histone H3 and total histone H3 levels were determined by Western blotting. B, H295R WT and ASAH1KD (pretreated with 5 μg/ml tet for 96 h) cells were crossed linked with 1% formaldehyde and the sheared chromatin immunoprecipitated with an antiacetyl histone H3 antibody. Acetyl-histone H3 levels at the proximal promoter regions of StAR, MC2R, CYP17A1, NR4A2, and DAX-1 genes were assessed by quantitative reverse chain reaction (qPCR). Purified DNA was quantified by real-time PCR and normalized to the ΔΔ cycle threshold values of input DNA. Data are expressed as fold change over untreated WT controls.