Effects of <i>BP-C1</i> on annexin V level on MCF-7 CELLS.

<p>Cells were treated with <i>BP-C1</i> (750 µg/ml) for 48 h and Flow cytometric analysis of annexin V-FITC/PI double-stained was performed. In each plot, the lower left quadrant (Q3) represents viable cells, the upper left quadrant (Q1) indicates necrotic cells, the lower right quadrant (Q4) denotes early apoptotic cells, and the upper right quadrant (Q2) represents necrotic or late apoptotic cells.</p

The effect of <i>BP-C1</i> on cell cycle progression.

<p>MCF-7 cells were treated with 750 µg/ml for 48 h. followed by treatment with RNase (100 µg/ml) and DNA staining with propidium iodide (PI). Cell cycle was analyzed using the FACSCalibur Cell Sorter.</p

Effects of <i>BP-C1</i> on annexin V level on T47D CELLS.

<p>Cells were treated with <i>BP-C1</i> (750 µg/ml) for 48 h and Flow cytometric analysis of annexin V-FITC/PI double-stained was performed. In each plot, the lower left quadrant (Q3) represents viable cells, the upper left quadrant (Q1) indicates necrotic cells, the lower right quadrant (Q4) denotes early apoptotic cells, and the upper right quadrant (Q2) represents necrotic or late apoptotic cells.</p

The effect of <i>BP-C1</i> on gene expression.

<p>MCF-7 cells were treated with 750 µg/ml for 48 h. After treatment, total RNA was isolated and reverse transcribed.Gene expression of pro-apoptotic genes was detected using the Applied Biosystems® TaqMan® Array Plates. HPRT1 and GAPDH genes were used as housekeeping genes for internal control to correct the potential variation in RNA loading.</p

Activation of caspase 8 (A) and caspase 9 (C) by <i>BP-C1</i>.

<p>Cells were treated with or without 750/ml of BP-C1 for 48 h. Cells were harvested and lysed. Activation of caspase-9 was determined by Western blotting as described under “Materials of Methods”. TA represents paclitaxel and 32c refers to Lx2-32c. Columns represent the densitometry analysis of the indicated proteins. The level of actin in the cells is represented in B and D.</p

Chemical structure of BP-C1 analog.

<p>R1 and R2 represent any kind of chain.</p

Dose - dependent growth inhibitory effect of <i>BP-C1</i> on MCF-7, T47D cells.

<p>Exponentially growing cells were incubated in the absence or presence of 100 to 1,000 µg/ml BP-C1 for 48 hours and the viable cells were counted as described under <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0085156#s4" target="_blank">Materials and Methods</a>. The results are presented as percentage of control and expressed as means ± standard deviation of three independent experiments in which each treatment was performed in triplicates.</p