FIG. 3.

(A and B) Production of IL-8 after stimulation of UROtsa (A) and A498 (B) cells with ATP-γ-S (100 μM), P2Y₁ agonist MRS2365 (0.1 and 1 μM), P2Y₂ and P2Y₄ agonist UTP (100 μM), or P2X agonist α,β-MeATP (100 μM) for 6 h. The amount of IL-8 produced by ATP-γ-S was set at 100%, and data are presented as means plus SEMs (error bars) (n = 3 or 5 for A498 cells and n = 5 or 7 for UROtsa cells). Statistical significance in panel A is indicated as follows: ***, P < 0.001 versus control or α,β-MeATP; †, P < 0.05 versus UTP; and *, P < 0.05 versus control or α,β-MeATP. Statistical significance in panel B is indicated as follows: ***, P < 0.001 versus control, MRS2365, UTP, or α,β-MeATP; and †, P < 0.001 versus control, MRS2365, or α,β-MeATP. (C and D) RT-PCR demonstrating expression of the P2Y₁ (327-bp), P2Y₂ (259-bp), and P2Y₁₁ (260-bp) receptor subtypes in UROtsa (C) and A498 (D) cells. Lanes M, molecular size markers (in base pairs). (E) Expression of P2Y₂ receptor transcript in A498 cells infected with the UPEC strain IA2. Representative results of three independent experiments are shown for RT-PCR. NS, nonstimulated cells.