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. 2010 Feb;77(2):288–297. doi: 10.1124/mol.109.059733

Fig. 10.

Fig. 10.

Complementation of TRH-stimulated receptor phosphorylation. HEK293 cells were cotransfected with DNA encoding two different TRH receptors, as shown; when only one receptor was expressed, a control plasmid was included. The 11Ala-TRH receptor was N-terminally tagged with HA, and all other receptors were tagged with V5 epitopes. Cells were incubated with or without 100 nM TRH for 5 min. Phosphorylated receptors were quantified by ELISA using antibody 6959. Signal from mock-transfected cells was subtracted from experimental values. The results are normalized to the phosphorylation of WT-TRH receptor in the same experiment. Coexpression of the 11Ala receptor increased surface expression of the WT and 6Q receptors by 21 and 31%, respectively. The increase in 6Q receptor expression does not account for the >500% increase in 6Q receptor phosphorylation caused by coexpression of 11Ala receptors. Shown are the mean ± S.E.M. values of three independent experiments performed in duplicate. ∗, P < 0.01.