Extended Data Fig. 5: Perturbing the SUMOylation site of AL3 affects its sub-cellular localisation. | Nature Plants

Extended Data Fig. 5: Perturbing the SUMOylation site of AL3 affects its sub-cellular localisation.

From: SUMO protease FUG1, histone reader AL3 and chromodomain protein LHP1 are integral to repeat expansion-induced gene silencing in Arabidopsis thaliana

Extended Data Fig. 5

a) A schematic of the Fluorescence Lifetime Imaging (FLIM) assay using transformed protoplasts. Scale bar = 5 μm b) Representative maximum intensity projections showing the localisation of GFP or GFP-AL3 or AL3 harbouring K178R mutation (GFP-AL3-K178R) in Bur-0 and fug1 mutant backgrounds 2.5 μm. c) Box and whisker plots of nuclear:cytoplasm ratio of GFP (blue, n = 6 in both Bur-0 and fug1) or GFP-AL3 (cyan, n = 16 in Bur-0 and n = 24 in fug1) or GFP-AL3-K178R (orange, n = 8 in Bur-0 and n = 14 in fug1) in Bur-0 and fug1 mutant backgrounds. Box plots show the 50th percentile and the whiskers show the max and minimum with the mean shown by a line in the graph. Each dot represents the quantification from an individual protoplast expressing the corresponding transgene. Statistical comparisons were done with a two sided Student’s t-test. P-values: **<0.01, ***<0.001, ****<0.0001. The first lane of panel b and the left half (Bur) of panel c are also used in the Main Fig. 3(h & i). The protoplast experiments were carried out at least twice with similar results.

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