Supplemental Figure-1 De Camilli

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Supplemental Figure 1 A No. at risk T T T

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Supplemental Figure-1 De Camilli Cyto+ATP+GTP Cyto+ATP+GTPγS Surface before incubation Surface after incubation % of Area Reduction

Supplemental Figure-2 De Camilli TfR-Phluorin 0min 10min Normalized intensity b PM-GFP 0min 10min Normalized intensity c 0min 10min Normalized intensity pixel

Supplemental Figure-3 De Camilli -FBP17 -dynamin dynamin FBP17 Intensity (a.u.) 5.1 5.4 5.7 6.3 6.9 7.8 9.0 μm -dynamin -endophilin endophilin dyn1 Intensity (a.u.) 6.6 7.2 7.5 8.1 8.4 8.7 9.0 μm -FBP17 -endophilin endophilin FBP17 Intensity (a.u.) Z (μm) 1 μm Away from surface Closer to surface

Supplemental Figure-4 De Camilli Cyto + ATP + GTPγS + Latru, 15min Cyto + ATP + GTPγS, 5min Cyto + ATP + GTPγS, 15min Number of the ccp Center Height of Individual Clathrin Coated Pits (nm)

Supplemental Figure-5 De Camilli FBP17 Actin b FBP17 Intensity (a.u.) + Latru + Latru + Latru Actin c Before t = 0 sec 10 sec 20 sec 30 sec 60 sec 300 sec FBP17 PM-GFP d PM-GFP Intensity (a.u.) FBP17

Supplemental Figure-6 De Camilli Protein A Sepharose Beads +anti-GFP antibody Protein A Sepharose Beads PM-GFP