N-cadherin is transported by VAMP8-positive transport vesicles. (A) Fluorescence images of SBP–EGFP–N-cadherin (Ncad) in the COS7 cells after biotin treatment. The arrowheads in the image at 30 min indicate vesicular structures transported from the Golgi to the PM. The arrows in the image at 60 min indicate PM-localized N-cadherin. Scale bar, 5 µm. (B–E) Fluorescence images of SBP–EGFP–Ncad (B and C) or SBP–EGFP–GPI (D and E) and mCherry–VAMP8 (B and D) or mCherry–VAMP2 (C and F) in the COS7 cells 60 min after biotin treatment. Blue arrowheads indicate EGFP and mCherry double-positive vesicular structures. Lower panels show magnified images of the boxes surrounded by dashed lines in the upper panels. Scale bars, 5 µm. (F) The rate of colocalization of SBP–EGFP–Ncad or SBP–EGFP–GPI and mCherry–VAMP8 or mCherry–VAMP2 (n = 5 images from each pair). (G) Time-lapse imaging of SBP–EGFP–Ncad and mCherry–VAMP8 after biotin treatment of the COS7 cells. The blue, red, white arrows indicate the same transported double-positive vesicular structures (see also Video 2). The box plots represent the maximum and minimum values. The centerlines of the box plots represent the medians of the data. Significance was calculated using two-tailed paired Student’s t tests. Statistical significance is indicated by ***P < 0.001.