Skip to main content
. 2011 Sep 1;12(5):427–435. doi: 10.4161/cbt.12.5.15984

Figure 2.

Figure 2

NTSR-1 proteins can be overexpressed and glycosylated in HEK293T cells. (A) HEK293T cells were seeded in 10 cm plates and allowed to attach before transfection with NTSR-1 constructs using lipofectamine 2000. After 48 h, the cells were lysed using RIPA buffer and samples containing equal total protein were analyzed using SDS gel electrophoresis. The blots were probed with anti-NTSR-1 antibody and quantified using Image-J software Representative plots from three separate experiments. (B) Transfected HEK293T cells were lysed using RIPA lysis buffer and NTSR-1 was immunoprecipitated using anti-NTSR-1 antibody. The protein was subjected to PGNaseF or O-type glycosidase and samples were analyzed using SDS-gel electrophoresis. Similar results were obtained when the immunoprecipitation experiment was performed on endogenously expressed NTSR-1 in MDA-MB-231 cells (data not shown); representative blots from three separate experiments.