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. 2001 Jul 31;98(16):9086–9091. doi: 10.1073/pnas.161290898

Figure 5.

Figure 5

ASGM1/mucin induction depends on a P2Y nucleotide receptor. (A) Involvement of a nucleotide receptor in the flagellin/mucin response demonstrated using the GPCR inhibitor suramin (250 μM, 45 min, in the presence of Triton X-100, 0.01%), the G-protein antagonist GDPβS (1 mM, 60 min, in the presence of Triton X-100, 0.01%), P2Y inhibitors [Reactive Blue-2 (100 μM, 45 min) and Acid Blue 129 (100 μM, 45 min)], and P2X inhibitors [PPADS (100 μM, 45 min) and NF023 (300 μM, 45 min)]. (B) Measurement of ATP in the extracellular medium of HM3 cells grown to 70% confluence and stimulated with 1:40 dilution of α-ASGM1 either alone or in the presence of 2.5 units/ml apyrase. (C) Luciferase reporter assay. HM3 MUC 2 cells pretreated with ATP permeability inhibitors, gadolinium (200 μM, 45 min), or glybenclamide (100 μM, 45 min) were then stimulated with α-ASGM1 antibody (1:100) for 4 h. Data are the mean ± SD values of triplicate determinations from three to six separate experiments.