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SleepyHollow02
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Untersuchte Arbeit:
Seite: 62, Zeilen: 1 ff. (komplett)
Quelle: Gu et al 2009
Seite(n): 0, Zeilen: 0
[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. These results indicate that actin redistribution is a mandatory step for the apoptotic response of mAR-stimulated colon cancer cells.

TAC: Testosterone-HSA Cyto B: cytochalasin B

Figure 15: Pro-apoptotic effects of testosterone-HSA, DHT and Estradiol in the absence or presence of inhibitors in Caco2 cells.

A) Quantitative APOPercentage apoptosis assay of testosterone-HSA stimulated Caco2 cells and similar experiments in the presence of cytochalasin B (Cyto B). Cells were exposed to 10-7 M testosterone-HSA for 24 hours and proapoptoric responses were assessed by the APOPercentage apoptosis assay. Equally, cells pre-treated or not with 10-7 M Cyto B or flutamide, were exposed to testosterone-HSA for 24 hours. Cells serum starved for comparable periods of time served as a positive control for apoptosis. Bars present the mean OD measured at 550 nm. ** P<0, 01, n=4.

B) Cells were pre-treated or not with Cyto B for 1h and then exposed or not to 10-7 M testosterone-HSA for 4h, lysed and incubated with the caspase-3 substrate DEVD conjugated to the chromophore pNA according to the manufacturer's instructions. Caspase-3 activity was measured at 405 nm. ** P<0, 01, n=4.

effects [30], the mAR-induced apoptotic response (Fig. 5A) and caspase-3 activation (Fig. 5B) were abolished. These results indicate that actin redistribution is a mandatory step for the apoptotic response of mAR-stimulated colon cancer cells.

Thus, we aimed to further evaluate the in vivo effects of albumin- PFrigou-arpeo p5totic effects of testosterone-HSA, DHT and estradiol in the absence or presence of inhibitors in Caco2 cells Pro-apoptotic effects of testosterone-HSA, DHT and estradiol in the absence or presence of inhibitors in Caco2 cells. (A) Quantitative APOPercentage apoptosis assay of testosterone-HSA, DHT or estradiol stimulated Caco2 cells and in the presence/absence of cytochalasin B (Cyto B) or flutamide. Cells were exposed to 10-7 M testosterone-HSA, DHT or estradiol for 24 hours and pro-apoptoric responses were assessed by the APOPercentage apoptosis assay. Equally, cells pretreated or not with 10-7 M Cyto B or flutamide, were exposed to testosterone-HSA for 24 hours. Cells serum starved for comparable periods of time served as a positive control for apoptosis. Bars present the mean OD measured at 550 nm (** P < 0.01, N = 4). (B) Cells were pre-treated or not with Cyto B or flutamide for 1 h and then exposed or not to 10-7 M testosterone- HSA for 4 h, lysed and incubated with the caspase-3 substrate DEVD conjugated to the chromophore pNA according to the manufacturer's instructions. Caspase-3 activity was measured at 405 nm (** P < 0.01, N = 4).

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