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>