The uncoupling phenotype is the consequence of an unbalance between PM tension and the force that the adaptor proteins can sustain. (A) Evolution of PM tension, monitored through the lifetime of the Flipper-TR probe measured by FLIM, upon TORC2 inhibition in TOR1-1 AVO3Δ^CT cells deleted or not of ENT1. Error bars represent the propagated error of mean values calculated from three independent experiments (with n > 10 cells). (B and C) Comparison of the evolution of the PM residency times of Sla1 (B) or Abp1 (C) upon TORC2 inhibition in TOR1-1 AVO3Δ^CT cells deleted or not of ENT1. Error bars represent SD of mean values calculated for n = 100 events from at least three independent experiments (****, P < 0.0001). (D) Comparison of the kinetics of appearance of the uncoupling phenotype upon TORC2 inhibition in TOR1-1 AVO3Δ^CT cells deleted or not of ENT1. Error bars represent SD of mean values calculated for n = 100 events from at least three independent experiments (*, P < 0.05; **, P < 0.01). n.s., not significant. (E) Deleting PM-binding N-terminal domain of Ent1 results in its colocalization with actin tails. Kymographs recorded across the PM showing Ent1(140–454)-GFP and Abp1-mCherry signals before and after 30 min of TORC2 inhibition. (F) Model illustrating that TORC2 inhibition induces an increase in PM tension that affects endocytosis at several levels.