cys-LT receptor expression and calcium signaling by LTD4 and
LTE4 in LAD2 cells. A, dose-dependent effects of
LTC4, LTD4, and LTE4 on the accumulation of
intracellular calcium in LAD2 cells. LAD2 cells were loaded with Fura-2-AM and
stimulated with the indicated concentrations of LTs in various orders.
B, effect of treatment with MK571 (1 μm, 5 min) on
calcium flux by Fura-2-AM-loaded LAD2 cells stimulated with the indicated LTs
(500 nm each). C, real-time PCR analysis of
CysLT1R, CysLT2R, and GPR17 transcripts expressed by
LAD2 cells. D, flow cytometry analysis of CysLT1R,
CysLT2R, and GPR17 proteins. CysLT1R and
CysLT2R were detected after permeabilization of LAD2 cells and
staining with anti-C terminus Abs. GPR17 staining was performed with an anti-N
terminus Ab both with (+perm) and without (–perm)
permeabilization. Shaded curves are staining with nonspecific rabbit
IgG. Results depicted are from single experiments, representative of at least
three performed for each assay.