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. Author manuscript; available in PMC: 2011 Oct 18.
Published in final edited form as: Nat Methods. 2011 Jul 10;8(8):691–696. doi: 10.1038/nmeth.1649

Figure 3.

Figure 3

Effect of acute proteasome inhibition on ubiquitin pools. (a,b) Ub-PSAQ analysis of lysates from HEK293 (a) and MEF (b) cell lines treated with either DMSO (vehicle) or the proteasome inhibitor MG-132 (1 μM) over 12 h. Error bars, ± s.d. (n = 3). *P < 0.05; **P < 0.01; and ***P < 0.005 (unpaired t-test). MonoUb, monoubiquitin conjugates. (c) Representation of ubiquitin pool components in HEK293 and MEF cell lines. (d) Ubiquitin western blots with antibodies FK2 and A100 showing ubiquitin-immunoreactive material for HEK293 cells treated with either DMSO (vehicle) or MG-132 (1 μM) over 12 h. Bars on top right of blots denote high-molecular-weight ubiquitin conjugates (HMW Ub conjugates), and ubiquitin-modified histone H2A (UH) and free ubiquitin are indicated. (e) Distribution of monoubiquitinated substrates in cytosolic and histone-enriched fractions from HEK293 cells treated with either DMSO or MG-132 (10 μM) for 6 h.