%0 Figure %A Fazzini, Alessandra %A D’Antongiovanni, Vanessa %A Giusti, Laura %A Da Valle, Ylenia %A Ciregia, Federica %A Piano, Ilaria %A Caputo, Antonella %A D’Ursi, Anna Maria %A Gargini, Claudia %A Lucacchini, Antonio %A Rosa Mazzoni, Maria %D 2014 %T PAR1 agonist-induced Gq but not Gi signaling is impaired in NCI-H28 cells. %U https://plos.figshare.com/articles/figure/_PAR_1_agonist_induced_G_q_but_not_G_i_signaling_is_impaired_in_NCI_H28_cells_/1227676 %R 10.1371/journal.pone.0111550.g004 %2 https://plos.figshare.com/ndownloader/files/1777556 %K plasma membrane localization %K mesothelioma cell lines %K dysfunctional PAR 1 %K Malignant Pleural Mesothelioma Cell Line %K mesothelioma REN cells %K PAR 1 %K cell surface PAR 1 expression %X

A, thrombin-induced intracellular Ca2+ mobilization in HMEC-1, Met-5A, and NCI-H28 cells. B, selective-PAR1-AP-induced intracellular Ca2+ mobilization in Met-5A and NCI-H28 cells. Serum and growth factor starved cells were loaded with Fluo-3AM to measure [Ca2+]i variations as indicated by changes in fluorescence intensity. Fluorescence was recorded before agonist addition (F0) and then every 3 seconds after thrombin (10 nM) or PAR1-AP (10 µM) addition for another 120 seconds. Data shown are mean ± SEM of a single experiment done in triplicate. Experiments were repeated two additional times with similar results. The results are reported as relative fluorescence (RF = F/F0 where F0 is basal fluorescence and F is fluorescence recorded after cell stimulation with the agonist). C, inhibition of isoproterenol stimulated cAMP production in Met-5A and NCI-H28 cells by different concentrations of thrombin in the presence and absence of 100 nM SCH 79797. D, no effect of the selective PAR1-AP on isoproterenol stimulated cAMP production in Met-5A and NCI-H28 cells. Serum and growth factor starved cells were exposed to different agonist concentrations. Assays were initiated by the addition of 1 µM isoproterenol. Production of cAMP was measured using a competition binding assay which includes the bovine adrenal cAMP binding protein and [3H]cAMP. Data shown are mean ± SEM of three independent experiments performed in triplicate. The differences between thrombin- and thrombin plus SCH 79797-treated cells were significant (**P≤0.01, ***P≤0.001) by one-way ANOVA followed by Bonferroni’s multiple comparison test (n = 3).

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