Fig. 3.

Ca²⁺ induces morphological, cell cycle, and bioenergetic changes in PWR-1E cells. a PWR-1E cells were cultured in KGM or KGM with 1.9 mM Ca²⁺ (i.e., PWR-1E high Ca²⁺) for 6 days. The cells were imaged using light and epifluorescence microscopy. For the epifluorescence images, the cells were stained with 40 nM DiOC₆(3) to visualize the mitochondrial network and 5 μg/ml of Hoechst 33342 to visualize the cell nucleus. The scale bars equal 18 μm. b Cell cycle distributions for the cells cultured as described in A were determined using hypotonic PI staining and flow cytometry as described in Fig. 1a. c Plots of O₂ consumption in PWR-1E cells cultured in KGM or KGM with 1.9 mM Ca²⁺ (i.e., PWR-1E high Ca²⁺) for 6 days as described in a. A media blank served as a negative control for O₂ consumption. The O₂ consumption results depicted in the inset graph are normalized for 10⁶ cells and expressed as the mean of triplicate samples for each treatment ± SD (error bars) (* P < 0.001 compared to the PWR-1E cells cultured under the normal low Ca²⁺ conditions)