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. Author manuscript; available in PMC: 2018 Jul 5.
Published in final edited form as: Toxicol Lett. 2017 May 18;276:85–91. doi: 10.1016/j.toxlet.2017.05.015

Figure 5. Precise regulation of Rpn4 protein levels in response to stress.

Figure 5

A) Steady-state Rpn4 protein levels after treatment with sodium arsenite (1 mM). The zero time point reflects Rpn4 induction after one hour of drug treatment. Sodium arsenite was then maintained in the culture or washed out, and Rpn4 levels were followed over time. Whole cell extracts were prepared and analyzed by SDS-PAGE followed by immunoblot with anti-Rpn4 antibody (upper panel) or anti-Pgk1 antibody (lower panel; loading control).

B) Steady-state Rpn4 protein levels after treatment with patulin (50 μg/mL). The zero time point reflects Rpn4 induction after one hour of drug treatment. Extracts were prepared and analyzed as in panel A.

C) Cycloheximide chase analysis of Rpn4 turnover. Rpn4 protein was induced by treatment with sodium arsenite (1 mM) for one hour. The drug was then washed out and cycloheximide (100 μg/mL) was added to inhibit new protein synthesis. Whole cell extracts were prepared and analyzed by SDS-PAGE followed by immunoblot with anti-Rpn4 antibody (upper panel) or anti-Rpn8 antibody (lower panel; loading control).