Cells expressing NTSR1 were stimulated with NTSR1 ligands. Phosphorylated ERK (pERK) and total ERK expression were assessed in whole cell lysates by Western blot analysis.
(A-B) Time course for NTS-stimulated pERK in HEK293T cells. NTSR1-expressing cells were treated with NTS in the presence or absence of the NTSR1/2 antagonist SR142948A. Controls cells lacking NTSR1 were stimulated only with NTS.
(C-D) Time course for SBI-553-stimulated pERK in HEK293T cells. NTSR1-expressing cells were treated with SBI-553 or vehicle.
(E-F) Time course for NTS and Vehicle or SBI-553 co-treatment-stimulated pERK in HEK293T cells. NTSR1-expressing cells were treated concurrently with NTS and vehicle or SBI-553.
(G-H) Time course for NTS and Vehicle or SBI-553 co-treatment-stimulated pERK in Gq-null cells. HEK293 cells genetically engineered to lack the Gq protein were treated concurrently with NTS and vehicle or SBI-553.
(I-J) Time course for NTS and Vehicle or SBI-553 co-treatment-stimulated pERK in β-arrestin1/2-null cells. HEK293 cells genetically engineered to lack β-arrestin1 and β-arrestin2 were treated concurrently with NTS and vehicle or SBI-553.
For details on statistical comparisons, see Table S3.