Excess centrosomes disrupt post-Golgi vesicle trafficking. (A) Fluorescence micrographs from time-lapse imaging depicting bleaching of Golgi edge (GalT::GFP) and subsequent recovery (FRAP) between TECs with one to two (top) and greater than two (bottom) centrosomes. Arrows, bleach time point; red circles, bleached areas; ROI, region of interest. Bars, 5 µm. see also Video 7. (B) One representative quantification of FRAP of Golgi marker between TECs with one to two or greater than two centrosomes from two experiments. (C) Box and whisker plots (middle bars, mean; boxes, top and bottom quartiles with error bars [SEMs]) of percentages of fluorescence recovery of Golgi marker 60 s after bleaching of TECs with one to two or greater than two centrosomes (n = 5 cells per group from two experiments). Statistics: Student’s t test; **, P < 0.01. (D) Representative single time point and time-compressed images (1–120 s) of mCherry-Rab6 vesicle trafficking time lapse in TECs with one to two or greater than two centrosomes. Insets are higher magnification of corresponding centrosomes (centrin::GFP). see also Video 8. Bars: (main images) 10 µm; (insets) 10 µm. (E) Mean percentage of vesicle trafficking to the dominant quadrant (see Materials and methods for details; NEC, n = 25; TEC 1–2, n = 17; TEC >2, n = 20 cells; three experiments). Statistics: Student’s t test; *, P < 0.05; **, P < 0.01. (F) Representative images of HUVECs with one to two or greater than two centrosomes expressing Rab6-mCherry. (left) Single slice is one video frame, and 1–60 s are time projections. Bar, 10 µm. (G) Mean percentage of vesicle traffic to the dominant quadrant in HUVECs with one to two or greater than two centrosomes (HUVEC 1–2, n = 10; HUVEC >2, n = 5; two experiments). Statistics: Student’s t test; means ± SEM; *, P < 0.05.