Physiological pH shift from 7.4 to 7.0 triggers a cytosolic Ca 2+ response that is propagated to mitochondria A) Representative fluorescence ratio traces of pH-dependent cytosolic Ca 2+ concentration ([Ca 2+ ] c ) changes monitored in untreated and PcTX1-pretreated WT neurons. Neurons were loaded with Fura-2-AM (1 μM) and pH change was carried out through superfusion. (B) Quantification of [Ca 2+ ] c increase as measured in (A) for untreated (n=3) and PcTX1-pretreated (n=3) neurons. (C) Representative fluorescence ratio traces of [Ca 2+ ] c changes induced by puffing with the pH 7.0 solution in WT and ASIC1a KO neurons loaded with Fura-2-AM. (D) Quantification of [Ca 2+ ] c increase as determined in (C) for WT (n=3) and ASIC1a KO (n=3) neurons. (E) Reresentative images of neurons (x10) that were loaded with Fura-2-AM before and after pH shift in the presence of Ca 2+ . The scale bars represent 100 μm. (F) Representative pH-dependent mitochondrial Ca 2+ concentration ([Ca 2+ ] m ) changes induced by superfusion and monitored by fluorescence in Rhod-2 AM (1 μM) loaded WT neurons untreated and pretreated with PcTX1. (G) Quantification of [Ca 2+ ] m changes as measured in (E) for untreated (n=7) and PcTX1-pretreated (n=7) neurons. (H) Representative fluorescence traces of [Ca 2+ ] m changes induced by puffing the pH 7.0 solution in WT and ASIC1a KO neurons. (I) Quantification of [Ca 2+ ] m changes determined as in (G) for WT (n=3) and ASIC1a KO (n=4) neurons. (J) Representative images of neurons (x20) that were loaded with Rhod-2 AM before and after pH shift in the presence of Ca 2+ . The scale bars represent 50 μm. (K) Representative fluorescence traces of pH-dependent [Ca 2+ ] m changes induced through superfusion in HEK293-T cells that overexpressed ASIC1a or were transfected with the empty vector (pcDNA). (L) Quantification [Ca 2+ ] m changes determined as in (I) for HEK293-T cells that overexpressed ASIC1a (n=3) and were transfected with pcDNA vector (n=3). (M) Rep
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