Effect of E2BSA and E2 on the modulation of apoptosis and autophagy.

<p>(<b>A</b>) Flow cytometry analysis of apoptosis in untreated and CMP treated SH-SY5Y cells in presence or absence of E2BSA, E2 and ERα antagonist MPP. Results obtained in a representative experiment are shown. Numbers in upper and bottom right quadrants of each plot refer to AV/PI double positive cells and to AV single positive cells, respectively. (<b>B</b>) Mean ± SD of the percentages of AV positive cells obtained in 10 independent experiments is reported. * <i>P</i><0.001. (<b>C</b>) Values of MFI <i>ratio</i> (Bcl-2/isotype control) are reported. The mean ± SD from 10 independent experiments is shown. * <i>P</i><0.01 for CMP treated cells <i>vs</i> all other conditions. Ctr, untreated cells. (<b>D</b>) Western blot analysis of LC3-II in untreated and CMP treated SH-SY5Y cells in presence or absence of E2 or E2BSA. Blots shown are representative of 10 independent experiments. (<b>E</b>) Densitometry analysis of LC3-II levels relative to β-actin is shown. Values are expressed as mean ± SD. * <i>P</i><0.01. Ctr, untreated cells. (<b>F</b>) Flow cytometry analysis of apoptosis in untreated and CMP treated SH-SY5Y cells in presence or absence of the autophagy inhibitor 3-MA and E2BSA. Mean ± SD of the percentages of AV positive cells obtained in 3 independent experiments is reported. (<b>G</b>) Western blot analysis of LC3-II in CMP treated SH-SY5Y cells in presence or absence of the autophagy inhibitor 3-MA and E2BSA. Blots shown are representative of 3 independent experiments.