USP25 reduces levels of ubiquitinated endogenous proteins associated with HRD1 or VCP/p97. Jessica R. Blount Aaron A. Burr Amanda Denuc Gemma Marfany Sokol V. Todi 10.1371/journal.pone.0036542.g005 https://plos.figshare.com/articles/figure/_USP25_reduces_levels_of_ubiquitinated_endogenous_proteins_associated_with_HRD1_or_VCP_p97_/311938 <p>A–C) HEK-293 cells were transfected as indicated. 48 hours later cells were harvested and constructs were immunopurified with bead-bound anti-V5 antibody to isolate V5-HRD1-associated protein (panels A and B), or with bead-bound anti-GFP antibody to isolate GFP-VCP-associated proteins (panel C). We did not detect ubiquitin species in western blots from stringent, denature/renature immunoprecipitations of HRD1 or VCP/p97 using RIPA buffer (data not shown), indicating that the ubiquitin signal we observe in this figure is due to species associated with HRD1 or VCP/p97. Shown are western blots of inputs from whole cells lysates and from immunopurified protein. Histograms are semi-quantifications of results from the left and other similar, independent experiments. Shown are means +/− standard deviations. P values for panels A and C are from Student-T tests.</p> 2013-02-20 05:26:58 levels ubiquitinated endogenous proteins hrd1