The translocation kinetics of His₆-LF_N through the (PA₆₃)₇ channel determined from macroscopic experiments compared with that determined from single-channel experiments. The protocol before stepping the voltage from 20 to 50 mV in the macroscopic experiments (the red and black curves) was the same as that described in the legend to Fig. 2. The cumulative probability distribution of translocation times (blue curve) was obtained from a survival curve (derived from single-channel experiments), such as that shown in Fig. 6, except that in this case the voltage was stepped from 20 to 50 mV (instead of to 48 mV). In the macroscopic experiment depicted by the red curve, the concentration of His₆-LF_N was so small that it only produced an 18% fall in conductance. Thus, virtually all of the blocked channels had only one His₆-LF_N bound to them. Note that the kinetics are identical to those obtained from the single-channel experiment (blue curve). In the macroscopic experiment depicted by the black curve, the concentration of His₆-LF_N was much larger, producing a 94% fall in conductance. In this case, ∼70% of the blocked channels had two or three His₆-LF_Ns bound to them (Fig. 4). Note that the rate of unblocking in this case was, as expected, slower than in the case when the blocked channels contained only one His₆-LF_N. The cartoon indicates the percentage of channels that have zero, one, two, and three His₆-LF_Ns bound to them when there was an 18% fall in conductance produced by His₆-LF_N (red numbers) and when there was a 94% fall in conductance (black numbers).