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. 2017 Nov 9;12(11):e0187481. doi: 10.1371/journal.pone.0187481

Fig 5. ecAT3.10 detects nucleotide-stimulated release of ATP from cells.

Fig 5

(A) A biphasic response was apparent on average when 100 μM ATP was added at time = 30 minutes to the static bath of ecAT3.10-expressing Neuro2A cells. The decrease in signal at time = 45 minutes is due to ARL67156-sensitive ectonucleotidase activity, and the subsequent increase in signal after time = 55 minutes reports a secondary release of endogenous ATP from cells. (B) Pretreatment with 100 μM ARL67156 abrogates the transient decrease in ectonucleotidase activity. (C) Addition of 100 μM ADP also stimulate a release of ATP from cells, which was potentiated by pretreatment with ARL67156 (D). Apyrase addition degrades extracellular ATP confirming a reversible ATP-specific sensor response. Solid lines show average responses from ecAT3.10, and dashed traces show average responses from the negative control ecATYEMK sensor (n = 3).