Figure 2. Expression and function of HA-H₁R in HEK293T cells.

(A) HEK293T cells were transfected with empty pEGZ (control) and pEGZ-HA-H₁R plasmids, and the expression of HA-H₁R was determined by Western blot using an anti-HA mAb. Data are representative of at least 3 independent experiments. (B) HEK293T cells were transfected as in A, fixed, permeabilized, and stained with an anti-HA mAb (red) and Topro-nuclear dye (blue). EGFP expression (green) represents transfected cells. Cells were visualized by confocal microscopy. Data are representative of at least 3 independent experiments. (C) HEK293 cells were transfected with pHA-H₁R-G_α11 fusion construct, and membrane fractions were isolated and used in [³⁵S]GTPγS binding assays in the absence (basal) or presence of 10^–4 M histamine. Samples were then used in immunoprecipitation using G_α11 antiserum, and the bound [³⁵S]GTPγS was measured by liquid-scintillation spectrometry. ***P < 0.001, Student’s t test.