Fig. 4: D-bodies are miRNA processing centres.
From: Phase separation of SERRATE drives dicing body assembly and promotes miRNA processing in Arabidopsis
a, Images (left) and quantification (right) of the distribution of pre-miR172b, miR172b/*, BFP–SE, DCL1–GFP and mCherry–HYL1 in pelleted droplet (P) and supernatant (S) before and after pre-miRNA cleavage. n = 3 independent experiments. P−Mg2+ versus +Mg2+for mCherry–HYL1 = 0.0110. b, Images (left) and quantification (right) showing the incorporation of BFP–SE, DCL1–GFP and mCherry–HYL1 in droplets without or with Mg2+. Scale bar, 5 μm. n = 10 fields of view examined over 3 independent experiments. P−Mg2+ versus +Mg2+ for BFP–SE = 6.1 × 10−11, P−Mg2+ versus +Mg2+ for DCL1–GFP = 3.3 × 10−14, P−Mg2+ versus +Mg2+ for mCherry–HYL1 = 1.1 × 10−13. c, Pull-down images (left) and quantification (right) showing the interactions of SE–Flag with DCL1–GFP and HYL1 without or with Mg2+. n = 3 independent experiments. P−Mg2+ versus +Mg2+ for mCherry–HYL1 = 0.0485. d, Pull-down images (left) and quantification (right) showing the interactions of SE–Flag with DCL1–GFP and HYL1 in the absence or presence of DCL1–GFP. n = 3 independent experiments. P−DCL–GFP versus +DCL1–GFP for HYL1 = 0.0429. e, Images (left) and quantification (right) showing the incorporation of mCherry–HYL1 into BFP–SE droplets in the presence of pre-miR172b or miR172b/*. Scale bar, 5 μm. n = 5 fields of view examined over 3 independent experiments. Pcontrol versus pre-miR172b = 7.3 × 10−9, Ppre-miR172b versus miR172b/* = 3.6 × 10−6. f, Pull-down images (left) and quantification (right) showing the interactions of SE–Flag with DCL1–GFP and HYL1 in the presence of pre-miR172b or miR172b/*. n = 3 independent experiments. Pcontrol versus pre-miR172b for DCL1–GFP = 0.0019, Ppre-miR172b versus miR172b/* for DCL1–GFP = 0.0094, Pcontrol versus pre-miR172b for HYL1 = 0.0310, Ppre-miR172b versus miR172b/* for HYL1 = 0.0147. All data are expressed as the mean ± s.d., and statistical significance values were determined using two-tailed Student’s t-test; *P ≤ 0.05. Uncropped gels and blots for a, c, d and f and statistical source data for a–f are provided in the source data.
