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0 (MicroCal, Northampton, MA, USA). The current traces were plotted with Origin software. Data are expressed as mean?��?standard error of the mean (SEM) of several cells (n) for each condition. Because of possible differences in cell size, membrane currents have been normalized and are shown as current densities. The statistical analyses were performed with two-tailed Student's t-test and values of P? http://www.selleckchem.com/products/BKM-120.html (Pasantes-Morales et?al., 1994). However, electrophysiological evidence for K+ channel activated by hypotonicity is still lacking. In order to unravel the contribution http://en.wikipedia.org/wiki/SWAP70 of the K+ channels to swelling-activated currents in astroglia, primary cultured rat cortical astrocytes were exposed to hypotonic solution in the absence and presence of the putative selective blocker of VRAC currents DCPIB (10?��M) (Nilius and Droogmans, 2003). Astrocytes were clamped at the holding potential (Vh) of ?40?mV, which is close to the zero-current potential under these experimental conditions (E0?=??34.6?��?2.4?mV, n?=?6), and a slow ramp stimulation protocol was applied from ?120?mV to +80?mV (inset to Figure?1A). Upon hypotonic challenge (��?=?60?mOsm��kg?1) a gradual increase in whole-cell currents within the entire range of membrane potentials and a positive shift in E0 to ?19.2?��?2.6?mV (n?=?6; P? http://www.selleckchem.com/products/BIBW2992.html were observed (Figure?1A). When DCPIB, at a concentration able to inhibit VRAC (10?��M) in cultured astrocytes (Abdullaev et?al., 2006), was added to the hypotonic solution a marked decrease in inward current was seen, indicating that in the negative range of membrane potentials the majority of hypotonicity-induced current was carried by VRACs (Benfenati et?al., 2009) (Figure?1A,B and Supporting Information Figure?S1A). Surprisingly, DCPIB caused an increase in outward current and the E0 shift to ?61.4?��?8.7?mV (n?=?6; P?