Survivin is stabilized and protects trophoblastic cells from apoptosis upon stress.

<p>(<b>A</b>) Western blot analysis. JAR cells were treated with 10 µM H₂O₂ and cellular lysates were prepared at 0, 4, 6, 8 and 10 h for Western blot analysis using antibodies as indicated. β-actin served as loading control. (<b>B</b>) Gene levels of survivin and Aurora B. Cells were treated as in (A) for 6 h and total RNA was isolated for real-time PCR analysis. The results are based on three independent experiments and presented as mean ± SEM. The RQ value of the untreated control cells is omitted and served just as a benchmark. (<b>C</b>) Immunofluorescence staining. JAR cells were treated with H₂O₂ for 6 h, fixed and stained for tubulin, survivin and DNA. Scale bar: 5 µm. (<b>D</b>) BeWo cells were treated with 20 µM forskolin for 0, 2, 4, 6, 8 and 10 h and cellular extracts were prepared for Western blot analysis with indicated antibodies. β-actin served as loading control. (<b>E</b>) Western blot analysis. JAR cells were non-treated (con), treated with scrambled siRNA (si con) or with siRNA targeting survivin (2.5, 7.5 and 10 nM) for 18 h and were then challenged with 10 µM H₂O₂ for additional 12 h. Cellular lysates were prepared for Western blot analysis with indicated antibodies. β-actin served as loading control. (<b>F</b>) Relative activity of caspase-3/7. The lysates from JAR cells treated as in (F) were used for evaluating caspase-3/7 activity. The results are presented as mean ± SD. (<b>G</b>) Western blot analysis. Primary trophoblasts were isolated from term placentas and stimulated with indicated concentrations of H₂O₂ for 6 h and cellular lysates were prepared for Western blot analyses with antibodies as indicated. β-actin served as loading control. (<b>H</b>) Western blot analysis. Isolated term primary trophoblasts were treated with increasing concentrations of forskolin for 8 h and cellular extracts were prepared for Western blot analyses with indicated antibodies. β-actin served as loading control.