Figure 4. The crowding effect of Ficoll... | Rockefeller University Press

$Figure 4. The crowding effect of Ficoll 400 on WT, G168D, and G113A MscS inactivation. (A–C) Pulse-step-pulse protocols were used to investigate the tension-driven inactivation of WT (A), G168D (B), and G113A MscS (C), with the difference between the initial and final pulse (black arrows) representing the relative amount of inactivation. After exposure to 10% Ficoll on the cytoplasmic side of the patch, WT MscS completely inactivates, G168D resists inactivation, and G113A displays an intermediate amount of inactivation (red arrows). (D–F) The same pulse-step-pulse protocol was applied at varying step pressures to several different patches of WT, G168D, and G113A (n = 3 for each) in the absence and presence of 5% Ficoll 400. The resulting fraction of available channels was plotted against tension to illustrate inactivation. The inactivation of WT MscS channels was greatly increased in the presence of Ficoll (D), whereas G168D remained largely unchanged (E) and G133A inactivated moderately (F). In the experiments requiring multiple stimulations (D–F), we used a lower concentration (5%) of Ficoll because 10% severely compromises the stability of patches.$

Figure 4.

The crowding effect of Ficoll 400 on WT, G168D, and G113A MscS inactivation. (A–C) Pulse-step-pulse protocols were used to investigate the tension-driven inactivation of WT (A), G168D (B), and G113A MscS (C), with the difference between the initial and final pulse (black arrows) representing the relative amount of inactivation. After exposure to 10% Ficoll on the cytoplasmic side of the patch, WT MscS completely inactivates, G168D resists inactivation, and G113A displays an intermediate amount of inactivation (red arrows). (D–F) The same pulse-step-pulse protocol was applied at varying step pressures to several different patches of WT, G168D, and G113A (n = 3 for each) in the absence and presence of 5% Ficoll 400. The resulting fraction of available channels was plotted against tension to illustrate inactivation. The inactivation of WT MscS channels was greatly increased in the presence of Ficoll (D), whereas G168D remained largely unchanged (E) and G133A inactivated moderately (F). In the experiments requiring multiple stimulations (D–F), we used a lower concentration (5%) of Ficoll because 10% severely compromises the stability of patches.

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