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Were covered with 1 ml of DMEM/F-12 containing 5 M Fura2/AM and incubated at 37 for 45 min. At the end of the period, the cover slips were washed twice with the physiological buffer solution containing (in mM): 125 NaCl, 5 KCl, 1.8 CaCl2, 2 MgCl2, 0.5 NaH2PO4, 5 NaHCO3, 10 HEPES, and 10 glucose, pH 7.4. The cells were incubated in physiological buffer for further 30 min to complete dye de-esterif