Figure 3
The modulatory action of Cl− applied to the cells maintained in 10 mM Cl− buffer. (A–C) The responses of cells expressing (A) mfT1R2a/mfT1R3, (B) zfT1R2a/zfT1R3, or (C) zfT1R2b/zfT1R3 with G16gust44. The cells were maintained in 10 mM Cl− buffer and stimulated with NaCl solution to obtain the final ionic composition, as shown in Supplementary Table S1, with or without corresponding agonists (200 µM L-Gln for mfT1R2a/mfT1R3, 160 µM L-Pro for zfT1R2a/zfT1R3, or 2 mM L-Ala for zfT1R2b/zfT1R3). The responses were plotted against the final [Cl−]o concentrations. (D–F) Na-gluconate solution was also applied to the cells expressing (D) mfT1R2a/mfT1R3, (E) zfT1R2a/zfT1R3, or (F) zfT1R2b/zfT1R3 to achieve the final ionic composition as shown in Supplementary Table S2. The responses to Na-gluconate solution were plotted along with the responses to the NaCl solution shown in (A)–(C) against the final osmotic pressure. Three independent experiments were conducted for each receptor, and representative results are shown as the mean ± SEM (n = 3 replicates). Significant differences were analyzed using a one-way analysis of variance (ANOVA) followed by Dunnett’s test (*p < 0.05, **p < 0.01 and ***p < 0.001 for vehicle vs NaCl + agonists; †p < 0.05 for vehicle vs NaCl; ‡p < 0.05, ‡‡p < 0.01 and ‡‡‡p < 0.001 for vehicle vs Na-gluconate + agonists). ∆RFU: Delta relative fluorescent units. VEH: responses against the 10 mM Cl− buffer with or without agonists.
