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. 2002 Jul;184(13):3740–3745. doi: 10.1128/JB.184.13.3740-3745.2002

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Copyright © 2002, American Society for Microbiology

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FIG. 3. — YopE_1-220-Npt, YopE_1-220-Ub-Npt, and YopE_1-220-Ub_Gly3,13-Npt are soluble proteins. Y. enterocolitica W22703 expressing either YopE_1-220-Npt, YopE_1-220-Ub-Npt, or YopE_1-220-Ub_Gly3,13-Npt was lysed in a French pressure cell. Crude extracts were subjected to ultracentrifugation at 100,000 × g. The soluble supernatant (So) and insoluble sediment (In) were separated, and proteins were precipitated with TCA and analyzed by immunoblotting (α-Npt or α-CAT). In the right panel, the signal intensities of Yop fusions in cell extracts are compared with the signal intensities in samples obtained by TCA precipitation of total cultures (T).