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Peptides were extracted from gels, transferred to a new tube and eluted as follows: 100 μl water was added to gels and samples were sonicated 10 min in a water bath followed by incubation with 5% formic acid in water, four extractions in 50% acetonitrile in 5% formic acid in water, one extraction in 70% acetonitrile and a final extraction in 100% acetonitrile