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NlpD binds PG in vitro. Ten micrograms of purified NlpD protein was incubated in binding buffer with or without PG for 2 h at 4°C. Supernatant was collected by centrifugation at 14,000 × g for 20 min at 4°C (lanes 1). Two wash steps followed by centrifugation (lanes 2 and 3) and the remaining pellet (lanes 4) were analyzed by SDS-PAGE followed by staining with Coomassie blue.
