TORC2 connects to endocytosis through two parallel pathways. (A) Evolution of PM tension, monitored through the lifetime of the Flipper-TR probe measured by FLIM, upon TORC2 inhibition by Rapamycin treatment in TOR1-1 AVO3ΔCT, TOR1-1 AVO3ΔCT FPK1Δ FPK2Δ, or TOR1-1 AVO3ΔCT ORM1Δ ORM2Δ cells. Error bars represent the propagated error of mean values calculated from three independent experiments (with n > 10 cells). (B and C) Evolution of the PM residency times of Sla1 (B) or Abp1 (C) upon TORC2 inhibition in TOR1-1 AVO3ΔCT FPK1Δ FPK2Δ cells. Error bars represent SD of mean values calculated for n = 50 events from at least three independent experiments (*, P < 0.05; ****, P < 0.0001; indicated in black when calculated to the WT at the same time point and in red when calculated to the initial time point of the FPK1Δ FPK2Δ cells). (D) Evolution of the percentage of cells displaying either an uncoupling phenotype or clustered endocytosis sites upon TORC2 inhibition in TOR1-1 AVO3ΔCT FPK1Δ FPK2Δ cells. Error bars represent SD of mean values calculated from three independent experiments (with n > 50 cells; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; indicated in black when calculated to the WT at the same time point and in red when calculated to the initial time point of the FPK1Δ FPK2Δ cells). Scale bars, 5 µm. n.s., not significant. Source data are provided in Table S1.
