Figure 3.
Cysteine 381,383 in NTSR-1 are required for optimal palmitoylation. HEK293T cells transfected with HA tagged WT-NTSR-1 and C381,383S NTSR-1 were serum-starved for 24 h before 4 h (3H) palmitate labeling. Cell lysates were subjected to immunoprecipitation using anti-HA antibody. One portion of the samples was analyzed using SDS-gel electrophoresis to determine equal expression level of NTSR-1 constructs. The other portion was analyzed using fluororadiography. The blots were sprayed with ENHANCE and exposed to X-ray film at −80°C for 4–6 weeks. Representative plots from three separate experiments. In this particular experiment, we used a HA tagged construct to increase the efficiency of receptor immunoprecipitation relative to using anti-NTSR-1. Mean ± SE, *p < 0.05.