VAEM micrographs of VAN3 and GNOM. (A–C) Double labeling of GNOM-GFP (A) and VAN3-mRFP (B) and merged image (C). GNOM-GFP and VAN3-mRFP colocalized at foci on the PM (arrowheads). (D–F) Double labeling of GNOM-GFP (D) and CLC-mKO (E) and merged image (F). GNOM-GFP and CLC-mKO colocalized at foci on the PM (arrowheads). (G–I) Double labeling of VAN3-GFP (G) and CLC-mKO (H) and merged image (I). VAN3-GFP and CLC-mKO colocalized at foci on the PM (arrowheads). (J) Colocalization efficiency of GNOM-GFP and VAN3-mRFP. (Left) Percentage of GNOM-GFP foci colocalized with VAN3-mRFP. (Right) Percentage of VAN3-mRFP foci colocalized with GNOM-GFP. As a negative control, the percentage of random colocalization (red frame is turned for 180°) is given. (K) Colocalization efficiency of GNOM-GFP and CLC-mKO. Left and right graphs give the percentage of GNOM-GFP foci colocalized with CLC-mKO and CLC-mKO foci colocalized with GNOM-GFP, respectively. As a negative control, the percentage of random colocalization (red frame is turned for 180°) is given. (L) Colocalization efficiency of VAN3-GFP and CLC-mKO. Left and right graphs give the percentage of VAN3-GFP foci colocalized with CLC-mKO and CLC-mKO foci colocalized VAN3-GFP, respectively. As a negative control, the percentage of random colocalization (red frame is turned for 180°) is given. (Scale bars in A–I: 1 μm.)