Fig. 3: CISD1 controls basal cytosolic calcium levels.

a, b Measurement of basal cytosolic calcium levels using FACS analysis. We stained PINK1 WT and KO (a) or Parkin WT and KO (b) MEF cells with Fluo-3 dye, and then measured the intensity of Fluo-3 fluorescence by FACS analysis. The cells were transfected with either control (gray) or CISD1 siRNA (blue). Bar graphs indicate relative basal calcium levels normalized to basal calcium levels of PINK1 WT (a) or Parkin WT (b) MEF cells. Three independent experiments were conducted and were quantified. Each sample was analyzed for 20,000 events using FACS. c Immunoblot analyses of cytosolic calcium signaling in PINK1 and Parkin WT or KO MEF cells when transfected with control (−) or CISD1 siRNA (+). Three independent experiments were conducted and the immunoblot band intensity for the phosphorylation of CaMKI and CaMKII were quantified (bar graphs in right). d Measurement of basal cytosolic calcium levels using FACS analysis in CISD1 WT (gray) or KO (blue) MEF cells. Bar graphs indicate relative basal calcium levels normalized to basal calcium levels of CISD1 WT MEF cells. Three independent experiments were conducted and were quantified. Each sample was analyzed for 20,000 events using FACS. e Immunoblot analyses of cytosolic calcium signaling in CISD1 WT or KO MEF cells. Three independent experiments were conducted and the immunoblot band intensity for the phosphorylation of CaMKI and CaMKII was quantitated (bar graphs in right). One-way ANOVA with Tukey’s multiple comparisons test was used (a–c), and two-tailed unpaired Student’s t test was used (d, e). ****p < 0.0001. ***p < 0.001. **p < 0.01. *p < 0.05. ns represents not significant. Source data and the exact p values are included within the Source Data file. All data are presented as mean ± SD.