Figure 4

Real-time quantitative PCR analysis of gene expression in HL-60 leukemic cells following sequential treatment with 5-AZA-CdR (A), DZNep (D), and/or TSA (T). The cells were treated with 20 nM 5-AZA-CdR and, at 24 h, 1,000 nM DZNep and/or 80 nM TSA were added to the medium. At 48 h the drugs were removed and at 72 h RNA was isolated for RT-PCR analysis as described in Methods. The relative gene expression is shown as mean ± SEM, n = 3. Statistical analysis for increase in relative expression of: CDKN1A (A), EGR3 (B), FBXO32 (C), CD86 (D), and SPARC (E): A + D + T > (A + D, A + T, T + D) P A<0.05 (one way ANOVA). EGR3 (B) A + D + T > A + D, n.s.