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

Figure 5.

Figure 5

Palmitoylation of glycosylated NTRS-1 isoforms is required for NTS-mediated MAP K mitogenic signaling. (A) HEK293T cells were seeded in 6 well plates and were then transfected with NTSR-1 constructs the next day. After 24 h, the cells were then serum-starved for another 24 h before 1 µM NTS stimulation for 15 min. The cell lysates were then analyzed using SDS-gel electrophoresis and blots were probed using P-ERK1/2 antibody. All blots were generated from the same gel and cropped in order to eliminate lanes irrelevant to this study. Representative blots from three separate experiments. Mean ± SE, *p < 0.05. (B) HEK293T cells were seeded in 6 well plates and were transfected with NTSR-1 constructs the next day. After 24 h, the cells were then serum-starved for another 24 h before 10 µM 2-BP treatment followed by 1 µM NTS stimulation for 15 min. The cell lysates were then analyzed using SDS-gel electrophoresis and blots were probed using a P-ERK1/2 antibody. Representative blots from three separate experiments. Mean ± SE, *p < 0.05. (C) MDA-MB-231 breast cancer cells were seeded in 6 well plates and serum starved for 48 h before 1 µM NTS stimulation for 15 min. The cells were lysed using RIPA buffer and equal total protein samples were analyzed using SDS gel electrophoresis. The blots were probed with P-ERK1/2 antibody and quantified using Image-J software. Representative blots from three separate experiments. Mean ± SE, *p < 0.05.