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. 2004 Oct 4;200(7):905–916. doi: 10.1084/jem.20040402

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Copyright © 2004, The Rockefeller University Press

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Figure 5. — Bim disappearance requires proteasomal activity. (A) Disappearance of Bim is prevented by a proteasome inhibitor. Hep2 cells were infected or not with C. trachomatis and treated with 500 nM MG132 and analyzed by Bim-specific Western blotting. (B) Infection with C. trachomatis does not increase proteasomal activity. As a measure of proteasomal activity, cleavage of Suc-LLVY-AMC was analyzed in cytosolic extracts from C. trachomatis and noninfected cells. ▪, uninfected; □, C. trachomatis–infected. The appearance of free AMC over time is shown. (C) Nonspecific proteasomal degradation of proteins in cytosolic extracts from infected cells. Cytosolic extracts of cells infected with C. trachomatis or not were incubated for 1.5 h at 37°C with GST-Bim_EL, GST-CED-4, and 500 nM proteasome inhibitor MG132 as indicated. Western blotting was performed using antibodies against Bim, tubulin, or GST.