FIGURE 5.

PKCδ or PKCϵ knockdown prevents NKCC1 internalization during PMA exposure. T84 cell monolayers grown to confluence on Transwell supports (6.5-mm insert diameter) were infected with shLacZ-, shPKCα-, shPKCδ-, or shPKCϵ-adenovirus (m.o.i. = 30) for 72 h. The cells were then exposed to vehicle (DMSO) or 100 nm PMA for 1 h. Subsequently, the cells were fixed in methanol, stained, and visualized by confocal microscopy. A, NKCC1 distribution detected by immunofluorescence with Alexa Fluor-488 (green) in shLacZ- cells treated with DMSO (left panel) or 100 nm PMA (right panel). The upper two panels represent X-Z sections and the lower two panels represent X-Y sections. B, same as A except the cells were transduced with shPKCα-adenovirus. C, same as A except the cells were transduced with shPKCδ-adenovirus. D, same as A except the cells were transduced with shPKCϵ-adenovirus. Scale bar, 10 μm.