Extended Data Fig. 7: COQ6DY does not impair calcium flux in AMs or BMDMs.
From: Novel coenzyme Q6 genetic variant increases susceptibility to pneumococcal disease
Cytosolic calcium was measured by confocal microscopy in Fluo-4 loaded BMDMs (a,b) and AMs (c,d) treated with pneumolysin. a. Peak amplitude of calcium signals from WT (grey) and DY (blue) BMDMs from a representative experiment of three independent experiments. b. Average peak calcium signal from n = 3 experiments. c. Peak amplitude of calcium signals from WT (grey) and DY (blue) AMs from three independent experiments (each experiment shown). d. Average peak calcium signal from n = 3 experiments. e. Representative kinetic calcium tracing from BMDMs stimulated with ionomycin (400nM). f. Peak amplitude of calcium signals from > 100 WT and DY BMDMs stimulated with ionomycin. g,h,i. Calcium flux measured by flow cytometry in Indo-1 loaded AMs. g. Representative kinetic calcium tracing from AMs stimulated with ionomycin (100nM) first in calcium-free media (arrow) to induce ER store release. 1mM CaCl2 was added at 120s as indicated by the darker blue bar to allow extracellular store-operated calcium entry (SOCE). h. Area under the curve (AUC) of ER store release (30–120s) and i. SOCE (150–450s) segments from n = 7 experiments. (a,c,f) Each symbol represents value from one cell, line at median, 95% CI shown. N=number of cells, given below each x-axis. (b,d,h,i) Each symbol represents average value for all cells analyzed in each experiment; bars show mean ± SD. N=number of experiments, given below x-axis. e) Each symbol shows mean ± SEM of all WT (gray) or DY (blue) cells analyzed in one representative experiment. Data were analyzed using unpaired Mann-Whitney (two-tailed) (a, c, h-i) or paired Wilcoxon rank test (two-tailed) (b, d).
