Skip to main content
. 2017 Oct 5;12(10):e0185992. doi: 10.1371/journal.pone.0185992

Fig 1. Identification of AP-2 subunits among MEX3C-associated proteins.

Fig 1

A. Identifying AP-2 α and β2 subunits among MEX3C-associated proteins. MEX3C interacting proteins were pulled down by anti-Flag antibody from cell lysates expressing MEX3C-2-Flag. M: molecular weight marker; kDa: kilodalton. B. AP-2 pulldown was MEX3C-dependent. Since affinity purified proteins were loaded, a loading control was unavailable. The lysates were prepared from equal numbers of HEK293 cells transfected with MEX3C-2-Flag expressing DNA or vector DNA, the purification was done in parallel, and equal volumes of eluents from MEX3C-Flag positive (+) and negative (-) lysates were loaded. C. RNase treatment did not eliminate MEX3C/AP-2 association. RNase A treatment eliminated MEX3C/THOC4 association, demonstrating that the RNase treatment worked. D. Comparisons of relative amounts of protein pulled down by MEX3C-2 with and without RNase treatment. Densitometry of protein bands was analyzed with Image J software. Protein expression was normalized by the respective MEX3C-2-Flag.