Effects of acute E
treatment on DAT protein levels in the membrane vs. intracellular compartment using the fixed cell immunoplate assay. After the cells had been serum-starved and NGFβ-treated for 2 days, 10 nM E2 or ethanol vehicle (ETOH) was added for the times indicated, before cell fixation and assay. Nonpermeabilizing conditions of fixation are shown in the left panel and permeabilizing conditions are shown in the right panel. ◇ represents clathrin Ab signal for monitoring the cell permeabilization status, as in previous figures. All E2-treated samples had DAT values significantly lower than ethanol-treated controls (p < 0.05).