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. 2014 Sep 16;24(10):1214–1230. doi: 10.1038/cr.2014.122

Figure 5.

Figure 5

BTZ stabilizes proteasome subunit, PSMC1, and proteasome assembly factor, PSMD10. (A) PSMC1 and PSMD10 were stabilized upon BTZ treatment (1.0 μM, 6 h). HEK293FT cells expressing PSMC1-mEGFPfu-mRFPf or PSMD10-mEGFPfu-mRFPf were treated with or without BTZ before cell lysis and immunoblotting analysis using anti-FLAG. Stabilities of each protein were assessed by comparing the abundances of the mEGFPfu fusions and the individual mRFPf references. (B) CHX chase analysis was performed with HEK293FT cells expressing PSMC1 or PSMD10 in the presence or absence of BTZ (1.0 μM) at the indicated time points. (C) After BTZ treatment (1.0 μM, 12 h), levels of endogenous PSMC1 or PSMD10 proteins in CZ-1 cells were assessed. (D) Levels of endogenous PSMC1 and PSMD10 proteins were higher in drug-resistant CZ-1/R cells than in CZ-1 cells. The expression levels of endogenous proteins were examined with immunoblotting analysis, using the indicated antibodies. (E) A schematic view of the feedback regulations of the proteasome homeostasis in responses to BTZ treatment.