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Fig. 2. BK promotes eNOS redistribution into a Triton X-100-insoluble fraction.
eNOS-GFP ECV 304 cells were incubated with BK (10 µM) or vehicle for 10
minutes then fractionated into cytosol and membrane fractions. Membrane
fractions were further fractionated into 0.5% Triton X-100-soluble (TS) and
-insoluble fractions (TIF). eNOS protein abundance was assessed in fractions
by western blot analysis. While BK stimulation did not influence the
distribution of eNOS in membrane (memb) and cytosolic (cytosol) fractions
(left panel), BK stimulation was associated with an enrichment of eNOS within
the TIF component of the membrane fraction (right panel). Control experiments
demonstrate that caveolin protein levels (cav) are enriched in membrane
fractions compared to cytosol, and TIF compared to TS (lower panel). Each
panel is representative of three separate experiments.