Figure 5. Pup-GGQ is not degraded by the Mtb proteasome in vitro.

A. Western analysis of the Pup-GGQ degradation assay: (Lanes 1-5) 0.1 µM Msm Mpa, 0.3 µM Opengate proteasome CP and 15 µM Pup-GGQ; (Lanes 6-10) 0.15 µM Msm Mpa, 0.2 µM WT proteasome CP and 15 µM Pup-GGQ. B. Activity assay for the Opengate and WT proteasome CPs. Both WT and Opengate proteasome CPs exhibit increased fluorescence after 1 h due to cleavage of the Suc-LLVT-AMC substrate indicating proteasome activity. C. Inhibition of WT proteasome CP activity by Bortezomib. Proteasome activity in the absence (black) and presence (gray) of Bortezomib. All experiments were performed in triplicate. The data are presented as mean +/- SEM. See also Figure S6 .