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. 2017 Sep 15;28(19):2479–2491. doi: 10.1091/mbc.E17-03-0162

FIGURE 2:

FIGURE 2:

Proteasome localizations in ubp6Δ and ubi4Δ cells. (A) Direct fluorescence microscopy of wild-type and mutant ubp6Δ cells expressing RP base Rpn1-GFP and Hta2-mCherry during cell proliferation and quiescence (top panels). Direct fluorescence microscopy of quiescent ubp6Δ cells expressing either Pre2-GFP or PR lid Rpn11-GFP and Hta2-mCherry (bottom panel). Scale bar: 3 µm. (B) Fluorescence of Pre2-, Rpn1-, and Rpn11-GFP in proliferating and quiescent wild-type and ubp6Δ cells was scored as nuclear (green), diffuse (gray), or in PSGs (red); n > 200 cells, three experiments, error bars show SD. (C) Direct fluorescence microscopy of quiescent ubi4Δ cells expressing Pre2-GFP. Scale bar: 3 µm. (D) Fluorescence of Pre2-, Rpn1-, and Rpn11-GFP in quiescent ubi4Δ cells was scored as nuclear (green), diffuse (gray), or in PSGs (red); n > 200 cells, three experiments, error bars show SD. (E) Levels of free ubiquitin were detected in cell lysates by SDS–PAGE and Western blot using antibodies against ubiquitin and GFP. Kar2 was probed as loading control.