Somatic Activation of Testis-Specific PDHA2 Gene Figure 4. DAC increases PDHA2 mRNA levels in SH-SY5Y cells. Real-time PCR analysis of PDHA2 steady-state mRNA transcript levels in SH- SY5Y cells treated with 5 mM DAC for the indicated time points (A), and with 5 mM DAC for 96 h and/or 0.25 mM TSA for 24 h (B). Values were normalized to the internal standard b-actin. Data represent means 6 SEM of at least three independent experiments and was expressed as pg of PDHA2 mRNA per ng of b-actin mRNA (* p,0.001; 1 p,0.01). doi:10.1371/journal.pone.0038076.g004 Figure 5. Recruitment of RNA pol II to the human PDHA2 gene. Chromatin from SH-SY5Y was prepared at 72 hours after treatment with 5 mM DAC and precipitated with antibodies directed against IgG, Sp1 and RNA pol II. After DNA recovery, the precipitates were evaluated by real-time PCR as described in Experimental Procedures. Results are expressed as fold change over IgG and represent means of at least three independent experiments 6 SEM (* p,0.001). doi:10.1371/journal.pone.0038076.g005 PLoS ONE | www.plosone.org 5 June 2012 | Volume 7 | Issue 6 | e38076