10.1371/journal.pone.0036542.g005 Jessica R. Blount Jessica R. Blount Aaron A. Burr Aaron A. Burr Amanda Denuc Amanda Denuc Gemma Marfany Gemma Marfany Sokol V. Todi Sokol V. Todi USP25 reduces levels of ubiquitinated endogenous proteins associated with HRD1 or VCP/p97. Public Library of Science 2013 levels ubiquitinated endogenous proteins hrd1 2013-02-20 05:26:58 Figure 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>