Extended Data Fig. 6: MEMB12 labels a heterogeneous network while NAG1, SYP31 and SYP32 label cisternae, related to Fig. 5.

(a, b) τ-STED morphometric analyses of either mCherry-MEMB12 x NAG1-EGFP (a) or α-MEMB immunostained compartments in NAG1-EGFP expressing plants (b). The circularity and solidity values were lower for α-MEMB than for mCherry-MEMB12 indicative that α-MEMB labels a network while mCherry-MEMB12 labels a tubulo-vesicular network. The binomial distribution of NAG1 values in Fig. 5j–t is explainable by the orientation of the cisterna during the acquisition, either from side or top view. Donut-like structures have an area, circularity and solidity index higher than rice grain structures (n = 176 NAG1-compartments including 88 “rice” and 88 “donuts” in a, n = 3 049 mCherry-MEMB12-compartments in a, n = 238 NAG1-compartments including 150 “rice” and 88 “donuts” in b, n = 1 837 α-MEMB -compartments in b). (c-h) τ-STED images of α-MEMB immunostained compartments (c, f) in mCherry-MEMB12 (d, g) expressing plants. Both α-MEMB and mCherry-MEMB12 label tubulo-vesiculated networks that share some overlap (e, h). (i) Quantification (n = 34 measures) of the co-localization between α-MEMB and mCherry-MEMB12 (e, h) using Pearson or Manders coefficient. As a control, the co-localization was calculated between α-MEMB and the NAG1-EGFP medial-Golgi. (j–r) τ-STED images of either NAG1-EGFP (j, m, p), EGFP-SYP32 expressed under the SYP32 promoter (k, n, q) or mRFP-SYP31 (l, o, r). (s, t) Morphometric analyses of j-r. SYP31, SYP32 and NAG1 label cisternae-like structures of different area, SYP31- or SYP32-compartments is smaller than the NAG1 medial-Golgi (out of 16 cells, n = 315 NAG1-compartments in s including 199 “rice” and 116 “donut” in t, n = 134 SYP32-compartments in s including 68 “rice” and 66 “donut” in t, n = 311 SYP31-compartments in s including 217 “rice” and 90 “donuts” in t. ^****P < 0.0001. In b, ^nsP = 0.9744 (NAG1-rice/α-MEMB). In s, ^nsP = 0.0849 (solidity). In t (area), ^**P = 0.0031 (NAG1-rice/SYP32-rice/SYP31-rice) and ^nsP = 0.5142 (NAG1-donut/SYP32-donut/SYP31-donut). In t (circularity), ^*P = 0.0305 (NAG1-rice/SYP32-rice/SYP31-rice) and ^**P = 0.0041 (NAG1-donut/SYP32-donut/SYP31-donut). In t (solidity), ^nsP = 0.5590 (NAG1-rice/SYP32-rice/SYP31-rice) and ^***P = 0.0003 (NAG1-donut/SYP32-donut/SYP31-donut)). Data are presented as violin plots with median represented as a line. For pair-wise comparison we used the non-parametric Mann-Whitney two-sided test. For multiple comparison in s and t, the Brown Forsythe test was used. Scale bars are 1 µm. Source numerical data are available in source data.

Source data