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. 2012 Oct 4;7(10):e46865. doi: 10.1371/journal.pone.0046865

Figure 5. DRSC26457 identifies dBest1 as Clswell.

Figure 5

(A & B) RNAi efficiently targeting dBest1 prevents significant I–induced H148Q-YFP suppression following hypo-osmotic stimulation. Fluorescence is in arbitrary units (a.u.). (A) Plate reader assay. DRSC26457 RNAi treatment resulted in a fluorescence decrease of 6.4% ±19. Control and DRSC16909 RNAi treatment resulted in fluorescence decreases of 40.5% ±9.1 and 43.7% ±6.2 respectively. * 240 mOSM NaCl and NaI fluorescence levels are significantly different (Student’s t-test, p<0.05). (B) Imaging assay. The fluorescence levels of individual S2R+ cells treated with control or dBest1 DRSC26457 RNAi were measured during hypo-osmotic stimulation in the sequential presence of Cl− and I−. The fluorescence of control cells decreased by 56% (n = 44); in contrast, the fluorescence of dBest1 DRSC26457 RNAi treated cells was suppressed by 15% (n = 174). (C) dBest1 DRSC26457 RNAi eliminated IClswell in S2R+ cells. Following dBest1 RNAi treatment I320 is not significantly different from I240 (Student’s t-test, p = 0.1). * control and RNAi treated I240 are significantly different (Student’s t-test, p = 0.02). (D) dBest1 W94C-gfp overexpression suppresses S2R+ IClswell. * control and W94C-gfp I200 mOSM are significantly different (Student’s t-test, p = 0.02). (E) Confocal images of dBest1 W94C-gfp overexpression in S2R+ cells. Images were obtained before (320 mOSM) and after swell (200 mOSM). Scale bar indicates 10 µm.