Figure 3. HRH4 activation induced growth arrest in gastric carcinoma cell lines.

(A) Mock-AGS and H4R-AGS cells were treated with 10⁻⁵ M histamine, CB or CB accompanied by HRH4 antagonist (JNJ7777120) pretreatment, and cell-cycle distributions were determined by propidium iodide flow cytometry analysis. Each value is the mean±s.e. of triplicate data representative for three independent experiments. *p<0.05 and **p<0.01 vs. Control, H4R-AGS cells without any treatment. (B) Colony-formation assay. 5×10³ Mock-AGS and H4R-AGS cells were treated as in Fig. 3A and maintained in G418 for 14 days, and the colonies were stained with Giemsa. The bar graph shows the absolute colony (≥50 cells) number±s.e. in duplicate experiments. *p<0.05 and **p<0.01 vs. Control, H4R-AGS cells without any treatment. (C) Mock-AGS and H4R-AGS cells were treated as in Fig. 3A, WST-1 (Roche) assay measuring the activity of mitochondrial dehydrogenases was performed following the manufacturer's instruction at 0-, 1-, 2-, 3- ,4- ,5- day time points. Each value is the means.e. of triplicate data representative for three independent experiments. Error bars represent standard deviation of the mean.