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Fig. 3. Centromere spatial localization in FKC8-activated hMSCs. Human MSCs at passage four were transduced with CenpA-GFP followed by FKC8 lentiviral vectors. Confocal images were taken from living cells before (mocked-treated) or 6 hours after 100 nM AP20187 treatment (+AP20187). Images were processed with TeloView. Bar, 5 µm. (A) 3D representation of a nucleus exhibiting a disk or a convoluted morphology. CenpA-GFP is shown in green. DIC image in the xy-axis reveals the nuclear rim. Underneath each image, a graph shows the spatial distance of centromeres from the center of mass (CM) after analyses in the algorithm centromeres. (B) Statistical analysis of centromere distribution in FKC8-transduced cells. The spatial localization of CenpA-GFP was evaluated using the centromeres algorithm. The average of CenpA-GFP dots close to center of mass (CM) (0-0.3 normalized distance) or close to the periphery (0.7-1 normalized distance) was calculated. FKC8-transduced cells were treated with 100 nM AP20187 (+AP) only or together with 50 nM Z-VAD.fmk (+AP+Z) for 5 hours. Control cells were mock-treated. AP20187-treated cells were sorted according to nuclear shape (round or convoluted). 50 nuclei were evaluated.
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