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. 2017 Jun 5;216(6):1715–1730. doi: 10.1083/jcb.201612123

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© 2017 Wang et al.

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Figure 7. — LysoK_VCa and its Ca²⁺ sensitivity regulate the refilling of lysosomal Ca²⁺ stores. (A and B) Acute application of paxilline (A) and quinidine (B) abolished the second ML-SA1-induced responses. Prolonged washout for 10–15 min led to a partial recovery of the responses. (C) Lysosome Ca²⁺ refilling in HEK-GCaMP3-ML1 cells treated with paxilline, quinidine, and IBTX. (D) Compared with WT MEFs, ML-SA1–stimulated refilled responses were reduced in GCaMP7-ML1-expressing KCNMA1 KO MEFs. (E) Lyso-SLO1^M513I/D898A currents at different concentrations of Ca²⁺ (0.1 and 10 µM). (F and G) Lysosomal refilling in GCaMP7-ML1–expressing KCNMA1 KO MEFs that were transfected WT SLO-mCherry and SLO1^M513I/D898A-mCherry. (H) GPN-induced refilled (the second) Ca²⁺ response, measured with Fura-2 imaging, was reduced in KCNMA1 KO MEFs. (I) Paxilline effects in lysosomal refilling in WT MEF cells. (J) Mean refilling responses in WT and KCNMA1 KO MEF cells. Statistical comparisons were made with variance analysis (Student’s t test). **, P < 0.01; ***, P < 0.001. Error bars indicate SEM.