Dual-color 4D super-resolution confocal live imaging microscopy (SCLIM) imaging of yeast cells expressing EGFP-Sed5 and Mnn9-mCherry (A–D), GFP-Vrg4 and Mnn9-mCherry (E–H), Sec21-2xEGFP and Mnn9-mCherry (I–L), and Mnn2-GFP and Mnn9-mCherry (M–P). (A, E, I, and M) Low-magnification images of the cells. The white broken lines indicate the edge of the cells. (B, F, J, and N) Time-lapse images of the single cisternae (white arrows in A, E, I, and M, respectively) in the cells. (C, G, K, and O) Time course changes in relative fluorescence intensities (F/Fpeak) of green and red channels in B, F, J, and N, respectively. (D, H, L, and P) Averaged time course changes in F/Fpeak of green and red channels (mean ± SEM). Time 0 was set as the midpoint between the green and red fluorescence peaks of each cisterna (n=13, 18, 17, and 6 cisternae for D, H, L, and P, respectively). Scale bars: 2 μm (A, E, I, and M) and 1 μm (B, F, J, and N).