10.1371/journal.pone.0101520.g006 Neelima P. Sidharthan Neelima P. Sidharthan Neville J. Butcher Neville J. Butcher Deanne J. Mitchell Deanne J. Mitchell Rodney F. Minchin Rodney F. Minchin Role of dimerization in SULT4A1 stability. Public Library of Science 2014 Biochemistry enzymology enzymes biocatalysis metabolism cell biology Molecular cell biology genetics gene expression toxicology Clinical medicine dimerization sult4a1 2014-07-02 03:04:23 Figure https://plos.figshare.com/articles/figure/_Role_of_dimerization_in_SULT4A1_stability_/1091914 <p>(A) IMR-32 cells transfected with wild-type (WT), TV>AE mutant or KTV>QAE mutant SULT4A1 were treated with 10 µg/ml cycloheximide and protein was collected at the different times. Western blots of protein were quantified by densitometry and normalized to tubulin as a loading control. Data are expressed relative to protein levels at time 0 (% Control). All results are mean ± s.e.m, n = 3. (B) Polyubiquitination of SULT4A1 wild-type and mutant proteins. IMR-32 cells were transfected with the different SULT4A1 FLAG-tagged constructs along with a HA-tagged ubiquitin construct. Following treatment with 20 µM MG132 for 8 hr, Western blots of HA-tagged proteins were performed. Expression of each construct was confirmed with anti-FLAG antibody (Input). Left panel represents immunopreciptation before protein denaturation while the right panel represents immunoprecipitation after protein denaturation. (C) Western blot for HA-ubiquitin on supernatants prior to immunoprecipitation.</p>