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Samples were washed twice with 1 X PBS, once with the CL-WS1 buffer [50 mM HEPES/KOH pH 7.5, 150 mM NaCl, 0.1% sodium deoxycholate, 1% Triton X-100, 1 mM EDTA, 1 mM PMSF, EDTA-free protease inhibitor cocktail], ground in liquid nitrogen, resuspended in 2 ml ice-cold CL-Lysis buffer [50 mM HEPES-KOH pH 7.5, 150 mM NaCl, 0.1% sodium deoxycholate, 1% Triton X-100, 0.1% SDS, 1 mM EDTA, 1 mM DTT, 1 mM PMSF, EDTA-free protease inhibitor cocktail], and sonicated using a Bioruptor UCD-200 at high mode for 15 cycles of 30 s on and 30 s off
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