After cells were treated with 1.2% isoflurane for 8 h, cytosolic calcium concentration ([Ca 2+ ] c ) was determined by flow cytometry. Briefly, the cells were harvested, stained with Fluo-3/AM and were then analyzed. (A-C) Representative results from one of six independent experiments on mutated β-amyloid precursor protein (APP)-transfected SH-SY5Y cells treated with (A) the control condition (Con), (B) xestospongin C plus isoflurane (Xes) and (C) isoflurane (Iso). (D-F) Representative results from vector-transfected SH-SY5Y cells treated with (D) Con, (E) Xes and (F) Iso. The positive rate of [Ca 2+ ] c loading with Fluo-3/AM was > 99%. (A-F) X-axis refers to the average value of calcium fluorescence intensity, and Y-axis refers to cell count. (G) Data are presented as the mean ± standard deviation of at least three separate experiments. *P