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Pak1 binding is sensitive to the spatial density of PIP2. Pak1 was incubated with 400 μm PC:PE (equimolar) liposomes containing increasing mole fractions of PIP2 (0–20%). Liposomes were sedimented and bound Pak1 was quantified by Western blotting. Results are shown for five replicate experiments (± S.D.) and the data were fit to a sigmoidal dose-response curve using GraphPad Prism to determine the Hill coefficient (nH).
