Canonical CCP closure. (A) A sequence of topographical HPICM images shows CCPs forming and closing with (red circles) and without (green circles) a cap on the same cell. (B) Topographical and fluorescent confocal image pairs show canonical CCP closure and associated Clc-EGFP fluorescence change. Clc-EGFP fluorescence remains at the pit’s location ∼180 s after pit closure. The green circle indicates CCP closure without membrane protrusion. (C) Depth increase during pit formation (n = 18). (D) Depth decrease before pit closure (n = 29). (C and D) Error bars represent SD. (E) Pit width distribution for pits that do not form caps (n = 133 from 26 independent experiments). (F) Lifetime distribution for pits that do not form caps (n = 65 from 26 independent experiments). (G) Pit width distribution for pits that form caps (n = 183 from 13 independent experiments). (H) Pit lifetime distribution for pits that form caps (n = 99 from 13 independent experiments). (C–H) All the data were measured using HPICM. (I) Pit closure without cap formation does not have associated Abp1-EGFP signal. White circles indicate the moment of CCP closure, as detected by ICM topography, and highlight the same area in the fluorescence confocal image where no Abp1-GFP fluorescence can be seen. (J) Pit closure without cap formation is associated with GFP-WT-DNM2 signal. White circles indicate the moment of CCP closure, as detected by ICM topography, and highlight the same area in the fluorescence confocal image where peak of DNM2-GFP intensity can be seen.