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Fig. 5 | Cell & Bioscience

Fig. 5

From: Proteomic characterization of endogenous substrates of mammalian ubiquitin ligase Hrd1

Fig. 5

The stability of OS9.1 and OS9.2 are differentially regulated. a, b Proteasome inhibition increases OS9.1 but not OS9.2. Lysates from cells treated with DMSO (Control) or MG132 (20 µm, 6 h) in triplicates were analyzed by immunoblotting (a). The graphs in b show the quantification (error bar, sem, n = 3). **p-value < 0.01. c, d Inhibition of p97 stabilizes OS9.1 but not OS9.2. Cells were treated with the indicated concentration of NMS-873 for 6 h followed by immunoblotting analysis. The graphs in d show quantification of 3 treatment experiments using 2 µm NMS-873. e Cycloheximide chase analysis of OS9 degradation in cells treated with MG132 and NMS-873. The graphs show quantification of 3 independent experiments. Error bars, sem, n = 3. f Chloroquine treatment stabilizes a fraction of OS9 in NP40 insoluble fraction. Immunoblotting analysis of NP40-soluble and insoluble fractions of cells treated with chloroquine (100 µM 16 h)

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