Figure 2.

NHERF1 inhibition of ERK1/2 does not require the PTH1R PDZ recognition domain. CHO-N10 cells were pretreated with 50 ng/ml Tet as indicated, and then they were transiently transfected with 1.0 μg of DNA/well of empty vector (pcDNA3.1⁺), wild-type PTH1R(ETVM), mutant PTH1R (ETVA), or truncated PTH1R(480stop). After 48 h, the cells were serum starved for 3–5 h, and then they were treated for 10 min with 1 nM PTH. The extent of phosphorylated and total ERK1/2, NHERF1, and PTH1R was determined as described in Materials and Methods. The results represent one of three independent experiments performed with similar results.