Fig. 2

Cx43 expressed on HUVECs has no effects on U937-HUVECs adhesion. a Gap 27 and 18-α-GA have no cytotoxic effects on HUVECs (n = 4). Cell vitality is detected using cell counting kit-8 kit assays. The data of absorbance are normalized to control. b Gap 27 and 18-α-GA have no effects on Cx43 expression on HUVECs (n = 4). Cx43 expression is detected by western blotting. The gray values of blots are normalized to control. c Gap 27 and 18-α-GA inhibit dye coupling between HUVECs (n = 5, *P < 0.05 vs control). The dye spread between HUVECs is detected by dye-coupling assay. The number of receiver cells around donor cell are normalized to control. d Gap 27 and 18-α-GA have no effects on U937-HUVEC adhesion. U937-HUVECs adhesion is detected by adhesion assays. The data of adhesion fraction are normalized to control. e ATP release from HUVECs is not changed when HUVECs are pre-treated with Gap 27 and 18-α-GA (n = 5). ATP release is detected with ATP bioluminescence assay kits. The intensity of bioluminescence is normalized to control. f The ADO content is not changed when HUVECs are pre-treated with Gap 27 and 18-α-GA (n = 5). The ADO content is detected using related ELISA kits. The data of absorbance are normalized to control. g Exogenous application of ATP and ADO reduces U937-HUVEC adhesion (n = 5, *P < 0.05 vs control); APCP reverses U937-HUVEC adhesion decrease provoked by exogenous ATP (n = 5, *P < 0.05 vs control; #P < 0.05 vs ATP group). U937-HUVECs adhesion is detected by adhesion assays. The data of adhesion fraction are normalized to control. Gap 27: 300 μM, for 1 h; 18-α-GA: 50 μM, for 1 h; APCP: 300 μM, for 1 h; exogenous ATP: 200 μM, for 1 h; exogenous ADO: 100 μM, for 1 h. ADO, adenosine; APCP, α, β-methylene ADP; HUVEC, human umbilical vein endothelial cell. All experiments are conducted in the presence of TNF-α. All data are presented as mean ± S.D. Multiple comparisons among groups are performed using repeated-measures one-way analyses of variance, followed by Tukey post hoc comparisons