Figure 6
MCU orchestrates Ca2+ entry into mitochondria of cells exposed to cl-CD95L. (a) BT549ShBclx cells reconstituted with BclxL-WT (BT549-BclxL-WT, upper panels) or BclxL-MT (BT549-BclxL-MT, lower panels) were loaded with a mitochondrial Ca2+ probe (Rhod2, red) and cell-permeable Mitotracker (green) to label mitochondria. Cells were pre-incubated for 1 h with a non-toxic amount of Ru360 (10 μM) and stimulated with cl-CD95L (100 ng/ml). Graphs depict the relative mitochondrial calcium concentrations in the indicated cells pre-treated with vehicle (left panel) or Ru360 (10 μM, right panel). (b) BT549ShBclx reconstituted with BclxL-WT and BclxL-MT cells were pre-incubated with Ru360 (10 μM) for 1 h and then stimulated with or without cl-CD95L (100 ng/ml). Cell migration was evaluated by the Boyden chamber assay. Migrating cells were fixed and stained with Giemsa. For each experiment, five images of random fields were acquired. To quantify cell migration, migrating cells were lysed and absorbance was measured at a wavelength of 560 nm. Values represent the mean±standard deviation of three independently performed experiments (**P<0.01, *P<0.05; two-way Mann-Whitney)
