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Samples were resuspended in 160 μL EPPS buffer (200 mM, pH 8.0) using a probe sonicator (1 × 10–15 pulses, power output 20%) and then digested with LysC (4 μL of 0.5 μg/μL per sample, resuspended in HPLC grade water, Wako-chemicals, Fujifilm #125-05061) for 2 h at 37 °C in a shaker incubator