Cdc42 colocalizes with ESCRTs in vivo. (A) Confocal spinning-disk still images of cells endogenously expressing Cdc42-mCherry^SW (magenta) and Chm7-GFP (green) in unbudded (top panel) or budded (lower panel) cells. Plot of normalized fluorescence intensity profiles of Chm7-GFP (green) and Cdc42-mCherry^SW (magenta) along yellow line shown in inset image. Cells that contain both Cdc42 spot and Chm7 spot where both are colocalized = 70% ± 0.71% (SD; n = 992; two trials). (B) Confocal spinning-disk stills of cells endogenously expressing Cdc42-mCherry^SW (magenta) and GFP-Snf7 (green) in unbudded (top panel) or budded (lower panel) cells. Plot of normalized fluorescence intensity profiles of GFP-Snf7 (green) and Cdc42-mCherry^SW (magenta) along yellow line shown in inset image. Cells that contain both Cdc42 spot and Snf7 punctae, where they are colocalized = 68% ± 0.64% (SD; n = 535; two trials). (C) Confocal spinning-disk stills of cells endogenously expressing Cdc42-mCherry^SW (red), GFP-HDEL (green), and mtagBFP2-Snf7 (blue) in G1 phase (top panel) or budding (lower panel). Plot of normalized fluorescence intensity profiles of Cdc42 (magenta), ER (green), and Snf7 (blue) along yellow line shown in inset image. All images are 2-µm maximum-intensity projections at 0.5-µm steps. All scale bars, 5 µm.