Fig. 4

ECT2/ECT3/ECT4 promote mRNA stability by recruiting PAB proteins. a Scatterplot showing the proteins bound to endogenous Arabidopsis ECT2 after RNase T1 treatment. The plot is based on the enrichment level (IP/control) and P-value. b BiFC assay showing the physical interaction between ECT2 and PAB2/PAB4 in Nicotiana benthamiana leaf cells. Scale bars = 20 μm. c Y2H assay showing the interaction of ECT2 with PAB2 and with PAB4 in yeast cells. d Correlation analysis of mRNA expression levels in Arabidopsis among ECT2 and PAB2, and PAB4 in the ATTED-II database (n > 10,000 samples; ρ, Spearman’s correlation coefficient). P-values were calculated with Pearson’s correlation analysis. e Top: predictions of PrLDs made by the “prion-like amino acid composition” (PLAAC; http://plaac.wi.mit.edu/); bottom: schematic diagram of ECT2 and its fragments. f Pull-down assay showing a direct interaction between PAB2 and PrLD domain of ECT2 in vitro. Purified MBP-PAB2 was incubated with GST-ECT2 fragments or GST alone, and pull-down assays were performed using GST magnetic beads, followed by immunoblot analysis with anti-GST and anti-MBP antibodies. g Overlapping of ECT2- and PAB2-binding targets in Arabidopsis. h The spatial distance distribution between the PAB2- and ECT2-binding sites. P-values were calculated using two-sided Mann–Whitney U test. i Cumulative distribution of relative mRNA half-life between ect2/3/4 and WT for Non-targets (black), ECT2 targets (blue), and ECT2 & PAB2 common targets (red). P-values were calculated using two-sided Mann–Whitney U test