TORC2 inhibition affects endocytosis at multiple stages. (A) Illustration explaining how the kymographs and time series were obtained. (B–E) Kymographs recorded either along (1) or across (2) the PM and time series of a single endocytic event (3) showing Abp1-mCherry signal and either Sla1-GFP (B and D) or Rvs167-GFP (C and E). They were recorded before (B and C) and after (D and E) TORC2 inhibition by 1-h Rapamycin treatment in TOR1-1 AVO3Δ^CT cells. The arrowhead is pointing at actin tails (D). (F–H) Evolution of the PM residency times of Sla1 (F), Abp1 (G), and Rvs167 (H), upon TORC2 inhibition in TOR1-1 AVO3Δ^CT cells. Error bars represent SD of mean values calculated for n = 50 events from at least three independent experiments (****, P < 0.0001; ***, P < 0.001; *, P < 0.05). n.s., not significant. (I) Evolution of Rvs167-GFP foci intensity and the percentage of cells displaying either an uncoupling phenotype or clustered endocytosis sites upon TORC2 inhibition in TOR1-1 AVO3Δ^CT cells. Error bars represent SD of mean values calculated from three independent experiments (with n > 100 cells or n > 150 foci; ****, P < 0.0001). Scale bars, 5 µm. Source data are provided in Table S1.