Fig. 1: PINK1 and Parkin activity modulate ER calcium release through IP3R calcium channel.
From: PINK1 and Parkin regulate IP3R-mediated ER calcium release
a, b Measurement of ER (a) and cytosolic (b) calcium modulation in WT (black) and Parkin KO (red) MEF cells. Similar experiments were also conducted for Parkin KO MEF cells expressing exogenous Parkin WT (blue), C431S (CS) mutant (green), or control empty vector (vector). 100 µM ATP was delivered to initiate IP3R-mediated calcium release. The right side bar graphs indicate the quantification of the normalized calcium traces using area-under-the-curve (AUC) of calcium release during ATP treatment. n = 162–231 cells. c, d Measurement of ER (c) and cytosolic (d) calcium modulation in WT (black) and PINK1 KO (red) MEF cells. Similar experiments were also conducted for PINK1 WT MEF cells expressing exogenous Parkin WT (blue) or control empty vector (vector), and PINK1 KO MEF cells expressing exogenous Parkin WT (green) or control empty vector (vector). The right side bar graphs indicate the quantification of the normalized calcium traces using AUC of calcium release during ATP treatment. n = 110–188 cells. e, f Measurement of IP3R activity. Cells were permeabilized for 100 sec with 20 µM β-escin in intracellular medium (ICM), and washed with ICM for 5 min. In all, 0.65 mM CaCl2 was then added to induce the influx of ER calcium. After a steady-state was achieved, 1 µM IP3 was introduced to evoke ER calcium release through IP3R. e Measurement of ER calcium release in WT (black, n = 72 cells) and Parkin KO (red, n = 68 cells) MEF cells. The bar graphs indicate the magnitude of the change during IP3 treatment. f Measurement of ER calcium release in WT (black, n = 65 cells) and PINK1 KO (red, n = 77 cells) MEF cells. Three independent experiments were conducted and were quantified (a–f). One-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test was used (a–d), and two-tailed unpaired Student’s t-test was used (e, f). ****p < 0.0001. ***p < 0.001. **p < 0.01. *p < 0.05. ns represents not significant. Source data, the exact p values, and n number of each experiment are included within the Source Data file. All data are presented as mean ± SD.
