Figure 8. DCA-p crosslinking is similar in channels favoring open or closed states.

Channels with β subunit V5-epitope tags and with furin sites mutated (α^Fβ^V5γ^F) to favor the closed state or with the degenerin site mutated (αβS518K^V5γ) to favor the open state were expressed in FRT cells. Cells were incubated with 50 μM DCA-p, exposed to UV light, following which surface proteins were labeled using a cell-impermeant biotin reagent. Samples were processed to isolate probe-labeled proteins at the cell surface. Input, surface and crosslinked surface fractions were subjected to SDS-PAGE/blot with anti-V5. Top, representative blot. Bottom, Quantified band densities were used to calculate relative total expression (B), surface expression (C), and crosslinked surface proteins. Individual data points (colored circles) and summary statistics (mean (SD), n = 10 culture wells) for are shown. Data were compared by Student’s t test.