Fig. 1: UBP5 is a nuclear protein that interacts with PRC2 and colocalises with PWO1.

a Localisation of UBP5 in the nucleus of N. benthamiana epidermal cells. Data represent results of three independent experiments, scale bar indicates 10 µm. b Transient and inducible expression of i35S::UBP5-GFP and i35S::PWO1-mCherry in N. benthamiana epidermal cells by co-transformation (i, i35S::UBP5-GFP; ii, i35S::PWO1-mCherry; iii, overlay). Arrows indicate speckles. Data represent results of three independent experiments, scale bar indicates 10 µm. c FRET-APB measurements for nuclei exemplified in b, with a distinction for speckle (spec) and non-speckle localisation. i35S::PWO1-GFP-mCherry measurements in speckles were used as positive control. The respective average efficiency per nuclei (n) is given in the plot. For box plot, the middle line represents the median; the upper and lower lines are the first and third quartile (Q1 and Q3); the whiskers indicate the upper and lower limits of data spread by subtracting 1.5* interquartile range (IQR) from Q1 and adding 1.5* IQR to Q3.Statistical significance was calculated using one-way ANOVA and the p-values are indicated in the figure. Error bars correspond to SE. d Y2H analyses confirm UBP5-PWO1 interaction and show UBP5-SWN and UBP5-EMF2 interactions. Yeast cells containing the different construct combinations on selective medium for plasmids (-WL; -Tryptophan, Leucine) or for reporter gene activation (-WLH; -Tryptophan, Leucine, Histidine). Serial dilutions were used. BD, GAL4-DNA binding fusion; AD, GAL4-DNA activation domain fusion. SWNΔSET, SWN construct lacking the SET domain. e Co-IP analyses confirming SWN-UBP5 interaction. IP was performed with anti-mCherry antibody and proteins were detected by western blot with anti-GFP. I, 5% input; IP, immunoprecipitation. Two biological replicates were performed.