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The LES is required for SusG surface exposure. WT SusG and the LES mutants were assessed for their surface exposure by proteinase K assays using whole cells (a) and OMVs (b). Proteinase K (PK) was added and incubated at 37°C; at different times (in minutes), aliquots were TCA precipitated and analyzed by SDS-PAGE and Western blotting using anti-SusG antibodies. We used 3×Flag-tagged BT_0766, a periplasmic soluble protein found in OMVs, as an outer membrane and OMV integrity control.
