10.1371/journal.pgen.1003275.g007 Martin Christmann Martin Christmann Tilo Schmaler Tilo Schmaler Colin Gordon Colin Gordon Xiaohua Huang Xiaohua Huang Özgür Bayram Özgür Bayram Josua Schinke Josua Schinke Sina Stumpf Sina Stumpf Wolfgang Dubiel Wolfgang Dubiel Gerhard H. Braus Gerhard H. Braus CSN targets DEN1/DenA for degradation in fungal and human cells. Public Library of Science 2013 targets degradation fungal 2013-02-07 00:47:22 Figure https://plos.figshare.com/articles/figure/_CSN_targets_DEN1_DenA_for_degradation_in_fungal_and_human_cells_/162842 <p>(A) Quantitative analysis of repeated western blot experiments displayed the differences in DenA abundance in fungal wild type and <i>ΔcsnG</i> cells. DenA levels of the different asexual developmental time points are shown relative to the vegetative (veg.) control for each strain. Anti-Actin was applied as loading control (statistics: 2-way ANOVA; n = 3; *p<0,05, **p<0,01). (B) Xpress-DEN1 was overexpressed in siGFP and siCSN1 human cells and steady state Xpress-DEN1 levels were estimated by western analysis with the α-Xpress antibody. Xpress-CSN1 was overexpressed in siCSN1 cells and DEN1 and CSN8 were probed with appropriate antibodies. (C) Xpress-DEN1 was overexpressed in siGFP human cells and the proteasome inhibitor MG132 was added 6 h before cell lysis at a final concentration of 10 µM. Cyclohexamide (CHX) was added in a final concentration of 10 µg/ml (D) Xpress-DEN1 was co-expressed in HeLa cells together with Xpress-CSN1wt, Xpress-CSN1(1–221) or Xpress-CSN1(222–527) in the absence or in the presence of MG132 (right panel), which was added 6 h before cell lysis. Cells were lyzed 24 h after co-transfection and lysates were analyzed by western blot using the α-DEN1 antibody (0 = only Xpress-DEN1).</p>