Figure 7.

DS-201 stabilized the membranous retention of BK_Ca channel proteins in HEK293 cells. (A) Schematic illustration showing the protocol for investigating BK_Ca channel internalization. Cell surface BK_Ca channels were first stained with anti-Flag antibody and then the α subunits were allowed to internalize for different time (0, 5, 10, 20, 40, and 60 min) at 37 °C. The remaining channels in the membrane were labeled by DyLight^®650-conjugated donkey anti-mouse IgG antibody and were detected by flow cytometry. (B) Normalized fluorescence intensity indicating the effects of DS-201 (80 μmol/L) on the membranous anchoring of BK_Ca channels (n=5). The time course of internalization was fitted by a single exponential equation: y=A1×exp(-x/t1)+y0. (C) Statistical results showing the normalized fluorescence intensity (FI) at 60 min of internalization (n=5). Note that DS-201 stabilized the membranous retention of BK_Ca channel α subunits. Mean±SEM. ^bP<0.05 vs control.