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Afterwards, cells were collected in HLB (50 mM HEPES-KOH, 150 mM NaCl, 1 mM EDTA, 0.1% sodium deoxycholate, 1% Triton X-100, 0.1% SDS, 1 mM PMSF, protease inhibitor cocktail, pH 7.5) and incubated on ice for 30 min