Appendix 3—figure 2. Pressure-dependent reduction of translocation rates.

(A) A solution of the free fluorophores Alexa488 and Alexa647 at a concentration of 100 nM was placed in the reservoir. The signal count rate was monitored at the exit of a pore with a diameter of 74 nm as a function of the applied pressure to the flow channel. Count rates were normalized to the values obtained in the absence of a pressure difference. Due to the pressure-induced hydrodynamic flow against the concentration gradient, the translocation rates decrease linearly with the applied pressure. (B) Event rates for Kap95 at a concentration of 1 µM acquired for three different open pores with diameters in the range of 67–84 nm. The event rates decrease markedly at a high pressure of 200 mbar, but remain approximately constant in the range below 100 mbar.