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Harvested cell pellets were lysed in PBST (1% Triton X-100 in PBS) and separated into soluble and insoluble fractions by centrifugation at 20,000 × g for 30 min at 4°C. The insoluble fraction was resuspended in a 2× sample buffer with an equal volume of soluble fraction and sonicated. Total lysates, soluble fractions, and insoluble fractions were resolved with SDS–PAGE and subjected to Western blot analysis.
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