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. 2009 Aug 4;284(42):28571–28578. doi: 10.1074/jbc.M109.010074

FIGURE 2.

FIGURE 2.

Activation of ICl,swell in HT29 cells requires TMEM16A channels. A, RT-PCR analysis indicates expression of TMEM16A, TMEM16F, TMEM16H, and TMEM16J in HT29 cells. M, marker; A–K, TMEM16A-K; Gs, Gl, short and long splice variants. B, original recordings of whole cell currents in HT29 cells, activated by a gradual increase of extracellular hypotonicity (17, 25, and 33%, arrows). Cells were voltage clamped in intervals from −50 to +50 mV. Partial replacement of extracellular Cl− by gluconate (open bar, 30 mm Cl−) inhibited whole cell outward currents. Treatment with siRNA for TMEM16A (si16A) reduced the swelling-activated whole cell current, when compared with cells treated with scrambled (scrbld) RNA. C, summary of whole cell conductance measured in HT29 cells under control conditions (open bars; normotonic bath solution) and after exposure to 33% hypotonicity (black bars). Mean ± S.E., (n) = number of cells measured. *, significant increase in whole cell conductance (paired t test). #, significant inhibition of ICl,swell by RNA interference knockdown of TMEM16A, when compared with treatment with scrambled RNA (unpaired t test).