Extended Data Fig. 5: Validation of the quantitative data obtained from the time-lapse airyscan acquisition of Fig. 4 and additional secGFP-FKBP-RUSH images.
From: ER-to-Golgi trafficking through a dynamic intermediate cis-Golgi tubular network in Arabidopsis
(a) Upper edge control: individual MEMB12-compartments could be tracked for an average track time of 22 sec. (b, c) Lower edge controls: one channel was flipped either horizontally or vertically, or rotated by 180°. (b) The number of ERES-associated MEMB12-compartments drastically decreased when one channel is rotated or flipped while the number of ERES-independent MEMB12-compartments is strongly increased. (c) The time of association between MEMB12 and the Sec16A-like MAG5 ERES marker strongly decreases when one channel is rotated or flipped. (d–g) Confocal images of root epidermal cells stably expressing both secGFP-FKBP-RUSH construct and mCherry-MEMB12 (repeated 3 times). (d) In absence of FKBP ligand, secGFP-FKBP-RUSH localizes to the ER. In presence of FKBP-ligand, sec-GFP localizes to small dotty structures as well as bigger pro-vacuole-like compartments after 10 min of incubation, or bigger vacuoles after 80 min of incubation. (h–j) Transient expression of the secGFP-FKBP-RUSH (h) in epidermal cotyledon cells stably expressing mCherry-MEMB12 (i, merged in j) (repeated 3 times). Vacuole-like compartments are labelled by secGFP-FKBP-RUSH upon incubation with FKBP-ligand. Data are presented as scatter plots with median represented as a line, n = 40 cells for all set of data. In b-c, ****P < 0.0001. All statistical tests were performed by non-parametric Mann-Whitney two-sided test. All scale bars are 4 µm. Source numerical data are available in source data.
