DNA methylation pattern on tumour suppressor genes in peripheral blood mononuclear cells of FA patients.

<p>PBMC from FA patients and healthy donor were isolated as in A, DNA was isolated and methylation profile was identified in 2 FA samples and 2 healthy donors samples using EpiTect Methyl II Complete PCR array (SABiosciences). Samples CTR1 and CTR2 represent control and FA1 and FA2 represent FA patients. Each row represents a tumour-supressor genes and each column represent a single DNA sample. The methylation degree are represented by the level of intensity of the square, red representing greater than 10% promoter hypermethylation, and green representing less than 10% promoter methylation (unmethylated) alleles for the tumour-suppressor gene.</p

DNA methylation pattern on tumour suppressor genes in peripheral blood mononuclear cells of FA patients.

<p>PBMC from FA patients and healthy donor were isolated as in A, DNA was isolated and methylation profile was identified in 2 FA samples and 2 healthy donors samples using EpiTect Methyl II Complete PCR array (SABiosciences). Samples CTR1 and CTR2 represent control and FA1 and FA2 represent FA patients. Each row represents a tumour-supressor genes and each column represent a single DNA sample. The methylation degree are represented by the level of intensity of the square, red representing greater than 10% promoter hypermethylation, and green representing less than 10% promoter methylation (unmethylated) alleles for the tumour-suppressor gene.</p

Effect of vorinostat on DEB-induced chromosome fragility of FA lymphocytes.

<p>(<b>A</b>) Lymphocytes metaphase without treatment. (<b>B</b>) Lymphocyte metaphase treated with DEB. (<b>C</b>) Lymphocyte metaphase treated with DEB after Vorinostat treatment. The red arrows indicate aberrant chromosomes characteristic of FA cells. (<b>D</b>) Number of breaks per cell after treatment with Vorinostat. (<b>E</b>) Number of breaks per cell after treatment with Vorinostat on DEB-induced breaks. The p values are indicated.</p

Number of breaks per cell in cultured lymphocytes from FA patients.

<p>The effect of the Vorinostat was calculated by the percentage of reduction of the number of breaks per cell in the Vorinostat treatments relatively to Spontaneous breaks and DEB.</p><p>Number of breaks per cell in cultured lymphocytes from FA patients.</p

Effect of vorinostat on epigenetic patterns on chromatin epigenetic modifications enzymes differentially expressed in FA mononuclear cells.

<p>PBMC from FA patients in culture were treated with 1μM vorinostat as indicated or vehicle (control) for 8h and gene expression quantified by qPCR.</p

FA patients have decreased expression of epigenetic chromatin modification enzymes.

<p>RNA was isolated from PBMC from 12 FA patients and 14 healthy controls and the expression of epigenetic regulator genes was quantified using the Human Epigenetic Chromatin Enzymes PCR array (SABiosciences). Each panel (A-L) represents the expression of the indicated genes in FA patients and control samples as described in the materials and methods section.(* 0.05 >p; ** 0.01>p; *** 0.001 > p). In each panel is indicated the values of FA1 and FA2 patients whose methylation profile was also determined.</p