Silencing TRADD reduces active caspase-8 (p18) levels and increases HeLa cell survival.

<p><b>Top panel: A</b>) The degree of TRADD knockdown was determined 48h after transfection with TRADD siRNA <b>(+)</b> or siRNA Negative Control <b>(</b>–<b>)</b> by quantitative RT-PCR. The TRADD mRNA levels results from 2 independent experiments (n = 8) are plotted relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and expressed as a fold change over the siRNA Negative Control. Error bars represent standard derivations. <b>B</b>) Following 48h after transfection with TRADD siRNA <b>(+)</b> or siRNA Negative Control <b>(-)</b> and 12h of treatment of HeLa cells with hesperidin (<b>h)</b> (2.5 μg/ml, <u>4.01 μM</u>), mangiferin (5 μg/ml, <u>11.84 μM) (<b>m</b>),</u> and hesperidin (2.5 μg/ml, <u>4.01 μM</u>) in a presence of mangiferin (5 μg/ml, <u>11.84 μM</u>) (<b>h+m</b>), the percentage of live cells were determined by trypan blue exclusion. The results were expressed as percentage of live control cells, which is represented as 100<i>%</i>. All the experiments were performed in triplicate at least two twice. Error bars represent standard derivations. Significant (p<0.05) changes are marked with an "*". <b>Bottom panel: C)</b> Following 48h after transfection with TRADD siRNA <b>(+)</b> or siRNA Negative Control <b>(</b>–<b>)</b> and 12h of treatment of HeLa cells with hesperidin (<b>h)</b> (2.5 μg/ml, <u>4.01 μM</u>), mangiferin (5 μg/ml, <u>11.84 μM) (<b>m</b>),</u> and hesperidin (2.5 μg/ml, <u>4.01 μM</u>) in a presence of mangiferin (5 μg/ml, <u>11.84 μM</u>) (<b>h+m</b>), the protein levels of procaspase-8 (55 kDa) and active caspase-8 p18 subunit (p18) were detected by Western Blot. Two individual samples (7<u> μ</u>g of total protein per lane) were tested for each treatment and the experiments were repeated three times. Protein levels were related to siRNA Negative Control Samples. Error bars represent standard derivations. Significant (p<0.05) changes are marked with an "*".