with10 Ryanodine, 10 Xestospongine C and 100 2-APB (see Methods). decrease in M M M Note, amplitude and influx of [Ca2+]C, in neurons transfected with siNCLX. Mean ± SEM of changes Right, in amplitude (peak (F- F0)/F0), and cytosolic Ca2+ influx rate (ΔF/s). nsiControl = 30; nsiNCLX = 25. ** p<0.05, ***p<0.001. Student's t-test. Supplementary Figure 2. siMCU impairs mitochondrial Ca2+ influx. A. Mitochondrial Ca2+ transients in HEK293T cells transfected with either siControl or siMCU following a puff application of 1 µM capsaicin (indicated by arrow). B. Mean ± SEM of mitochondrial Ca2+ influx and efflux rate (ΔF/s) in siControl (black, and siMCU (red, ** p < 0.01; Student's t-test. n=120) n=95). Supplementary Figure 3. Li+ regulates the capsaicin-dependent cytosolic and mitochondrial Ca2+ transients. A. representative traces of changes in fluorescence of HEK293T cells loaded Left, with Fura 2AM, co-transfected with TRPV1 and either siControl or siNCLX, following a short application of 1 µM capsaicin (indicated by arrow) in the presence of Na+ or Li+ in bath solution. mean ± SEM of peak ratio (peak (F- F0)/F0) and capsaicin-induced cytosolic Ca2+ influx rates Right, for siControl and siNCLX (n=110). B. representative traces of changes in mitochondrial Ca2+ Left,
with10 M Ryanodine, 10 M Xestospongine C and 100 M 2-APB (see Methods). Note, decrease in amplitude and influx of [Ca2+]C, in neurons transfected with siNCLX. Right, Mean ± SEM of changes in amplitude (peak (F- F0)/F0), and cytosolic Ca2+ influx rate (ΔF/s). nsiControl = 30; nsiNCLX = 25. ** p<0.05, ***p<0.001. Student's t-test. Supplementary Figure 2. siMCU impairs mitochondrial Ca2+ influx. A. Mitochondrial Ca2+ transients in HEK293T cells transfected with either siControl or siMCU following a puff application of 1 µM capsaicin (indicated by arrow). B. Mean ± SEM of mitochondrial Ca2+ influx and efflux rate (ΔF/s) in siControl (black, n=120) and siMCU (red, n=95). ** p < 0.01; Student's t-test. Supplementary Figure 3. Li+ regulates the capsaicin-dependent cytosolic and mitochondrial Ca2+ transients. A. Left, representative traces of changes in fluorescence of HEK293T cells loaded with Fura 2AM, co-transfected with TRPV1 and either siControl or siNCLX, following a short application of 1 µM capsaicin (indicated by arrow) in the presence of Na+ or Li+ in bath solution. Right, mean ± SEM of peak ratio (peak (F- F0)/F0) and capsaicin-induced cytosolic Ca2+ influx rates for siControl and siNCLX (n=110). B. Left, representative traces of changes in mitochondrial Ca2+
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