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. Author manuscript; available in PMC: 2016 Jan 19.
Published in final edited form as: Chembiochem. 2014 Dec 4;16(2):284–292. doi: 10.1002/cbic.201402491

Figure 4.

Figure 4

UKP13-(PEG)494-6Ahx-PR825 (probe 4) crosslinks multiple pairs of catalytic subunits. A) The purified 20S immunoproteasome (IP) was treated with DMSO, epoxomicin (Epx), or probe 4 prior to immunoblotting for β1i, β5i, or β2i. B) The purified 20S immunoproteasome was treated with DMSO or epoxomicin, or pretreated with DMSO or the indicated subunit-selective inhibitors prior to treatment with probe 4 as indicated. β5i was detected by immunoblotting. C) The purified 20S constitutive proteasome (CP) was treated with DMSO, YU-102, epoxomicin, or probe 4 prior to immunoblotting for β1 or β5. D) The purified 20S constitutive proteasome was treated with DMSO or YU-102, or pretreated with DMSO or the indicated subunit-selective inhibitors prior to treatment with probe 4 as indicated. β1 was detected by immunoblotting. Irrelevant lanes were removed from blots shown in A, Cand D. CL = crosslinked subunit.