How intracellular fluid flows influence the formation of complex patterns NatureComms
and S4. Sample channels were coated with lipid bilayers composed of DOPC:DOPG in a molar ratio 67:33 substituted with 0.01–0.02% mol TopFluor Cardiolipin, where the latter allowed us to confirm full bilayer formation before the experiment. SUVs for lipid bilayer formation were prepared via swelling followed by stepwise extrusion with final pore size of 40 nm. Before the experiment, chambers were filled with SUV solution, incubated for 1 h at 37 °C, then rinsed thoroughly with Min buffer .
To check whether protein concentrations changed due to sticking to the comparably large internal surfaces of the tubing, we collected a portion of the original solution as well as the sample extracted from the closed-circle system at the end of every experiment and ran them side-by-side on a gel to calculate an estimate for the loss, which typically was ~30% for MinD and ~50% for MinE. In the experiments described, the corrected ratios are given along with the initial ratios.
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