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Quelle: Gu et al 2009
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Figure 14: Modulation of the dynamic equilibrium rapid tubulin reorganization in testosterone-HSA stimulated Caco2 cells.

Caco2 cells treated or not with 10-7 M testosterone-HSA for different time points were cultured in coverslips, fixed and stained with rabbit anti-α-tubulin. Anti-rabbit-FITC was used as secondary antibody and DRAQ5™ for nuclei staining. Confocal laser scanning microscopy analyzed samples. Magnification, ×100.

Previously, it has been reported that activation of mAR with non permeable testosterone derivatives induced pro-apoptotic responses [Gu S Diploma Thesis, Gu S, et al. 2009]. However, the mechanism regulating the mAR-induced apoptotic responces is still unknown. In recent years, the cross-talk between actin cytoskeleton components and apoptotic signaling has attracted specific interest. Indeed, modifications of actin dynamics seem to be crucial for apoptotic responses [Gourlay CW, et al. 2005, Franklin-Tong VE,et al. 2008]. More recently the functional role of actin reorganization in regulating the pro-apoptotic responses induced by mAR was established in prostate cancer cells [Papadopoulou N, et al. 2008,] Based on these results we assessed the mAR-dependent apoptosis and caspase-3 activation in the presence of anti-actin drugs. As shown in Figures 15A,B, in Caco2 cells pre-[treated with cytochalasin B, at a concentration (10-7M) which blocks actin redistribution without exerting toxic effects [Stournaras et al 1996], the mAR-induced apoptotic response (Fig 15A) and caspase-3 activation (Fig 15B) were abolished.]

To establish the functional role of actin reorganization in regulating the pro-apoptotic responses induced by mAR, as previously reported for various cell systems [8,28,29], we assessed mAR-dependent apoptosis and caspase-3 activation in the presence of antiactin drugs. As shown in Fig. 5A, B, in Caco2 cells pretreated with cytochalasin B, at a concentration (10-7M) which blocks actin redistribution without exerting toxic PFrigou-arpeo p4totic effects of testosterone-HSA in Caco2 cells Pro-apoptotic effects of testosterone-HSA in Caco2 cells.

Rapid tubulin reorganization in testosterone-HSA stimulated Caco2 cells. Caco2 cells treated or not with 10-7 M testosterone-HSA for different time points were cultured in coverslips, fixed and stained with rabbit anti- -tubulin. Anti-rabbit-FITC was used as secondary antibody and DRAQ5™ for nuclei staining. Confocal laser scanning microscopy analyzed samples. Magnification, ×100.

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