Figure 1 |. 2-Bromodifluoroacetylamino-5,6,7,8-tetrahydro-4H-cyclohepta[b]thiophene-3-carboxylic acid o-tolylamide (TM_inh-23) pretreatment blocks Ca⁺²-activated Cl⁻ secretion in Fischer rat thyroid (FRT) cells and rat primary vascular smooth muscle cells.

(a) Chemical structure of TM_inh-23. (b) FRT cells expressing transmembrane member 16A or anoctamin-1 (TMEM16A) were pretreated with indicated concentrations of TM_inh-23 for 10 minutes before the addition of ionomycin (1 μM). Original data are shown on the left, and averaged data (mean ± SEM; n = 4) are shown on the right. (c) (Left) Rat pulmonary artery smooth muscle cells (RPASMCs) transduced with lentivirus to express halide-sensing yellow fluorescent protein (YFP; bar = 50 μm). (Middle) YFP fluorescence quenching in response to the extracellular addition of iodide, with or without adenosine triphosphate (ATP) and 3 μM TM_inh-23. (Right) Summary of the percentage inhibition of I- influx by TM_inh-23 in RPASMCs (mean ± SEM; n = 11–14). (d) Cell viability assayed by Alamar blue in RPASMCs incubated for 24 hours with vehicle control (0.1% dimethylsulfoxide [DMSO]) or 3 μM TM_inh-23. The 20% DMSO was used as positive control. Data are given as mean ± SEM, n = 6 wells per group, 1-way analysis of variance and post hoc Newman-Keuls multiple comparisons test. ***P < 0.001, NS, not significant. To optimize viewing of this image, please see the online version of this article at www.kidney-international.org.