Figure 6

qPCR validation of house-keeping and cell-line specific HDACi-induced gene expression changes. HCT116 and HT29 cells were treated with 2 μM vorinostat or 50 nM LBH589 for 6, 12 and 24 h. Total RNA was extracted and qPCR analysis was performed as described in the 'materials and methods' using the primer sets given in Table 4. Histogram bars represent the mean ± SD for two independent RNA isolations analyzed in triplicate. (A) Verification of unaffected 18s and GAPDH expression with HDACi treatment. GAPDH was normalized to 18s and 18s was normalized to GAPDH. qPCR validation of the induction of (B) THBS-1, (C) AVEN (D) AURKB (E) HIST1H1C. All genes were normalized to GAPDH, * denotes a p-value < 0.05 for both HDACi treatment groups when compared to respective time-matched control.