Portland Press

Figure 2

Panel (A) shows the effects of 4 h pre-treatment with the Ca2+ chelator BAPTA-am (10 µM) upon cell viability of Cos-7 cells that were then exposed to DOX (7 µM) for 24 h. Data are presented as the mean ± SD of four replicates. (B) Cells were transfected with GFP-LC3 for 48 h and then treated with 7 µM DOX for 4 h prior to analysis by fluorescence microscopy. Inset shows a cell with clear puncta indictive of autophagosome formation. The experiment was repeated in triplicate and SD determined of fluorescent cells that clearly showed the presence of puncta either in the presence or absence of DOX. Statistical analysis via two-tailed Student’s T-test was performed in comparing cell viability levels in DOX-treated cells in the presence and absence of BAPTA-am, or % cells showing GFP-LC3 puncta when treated in the absence or presence of DOX. Probability values **P<0.01, ***P<0.005.

Doxorubicin causes Ca²⁺-dependent cell death and autophagosome formation

Panel (A) shows the effects of 4 h pre-treatment with the Ca²⁺ chelator BAPTA-am (10 µM) upon cell viability of Cos-7 cells that were then exposed to DOX (7 µM) for 24 h. Data are presented as the mean ± SD of four replicates. (B) Cells were transfected with GFP-LC3 for 48 h and then treated with 7 µM DOX for 4 h prior to analysis by fluorescence microscopy. Inset shows a cell with clear puncta indictive of autophagosome formation. The experiment was repeated in triplicate and SD determined of fluorescent cells that clearly showed the presence of puncta either in the presence or absence of DOX. Statistical analysis via two-tailed Student’s T-test was performed in comparing cell viability levels in DOX-treated cells in the presence and absence of BAPTA-am, or % cells showing GFP-LC3 puncta when treated in the absence or presence of DOX. Probability values **P<0.01, ***P<0.005.

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