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The cells were subsequently washed with 250 l lysis buffer, incubated for 15 min with 500 l lysis buffer, and incubated for 10 min in 500 l stimulation buffer (similar to lysis buffer but with 0 or 5 mmol/l ATP and 1.65 or 6.65 mmol/l CaCl 2 ) at 37 C, 5% CO 2