Extended Data Fig. 5: Ell3 KD affects spindle assembly.
From: Maternal ELL3 loss-of-function leads to oocyte aneuploidy and early miscarriage
a, 3 h after GVBD, control and Ell3 KD mouse oocytes were immunostained with anti-BubR1 and anti-centromere (CREST) antibodies. Scale bar, 10 μm. b, BubR1 intensity in the oocytes depicted in a, for both the control (n = 24) and Ell3 KD (n = 23) groups. c, Control and Ell3 KD mouse oocytes at the MetI stage were immunostained with anti-BubR1 and CREST antibodies. Scale bar, 10 μm. d, BubR1 positive percentage of the oocytes depicted in c, for both the control (n = 19) and Ell3 KD (n = 23) groups. e, Representative images showing spindles and chromosomes in control, Ell3 KD and human ELL3-rescued mouse oocytes. Green, microtubules; blue, DNA. Scale bar, 10 μm. f, MI spindle abnormality rate in control (n = 131), Ell3 KD (n = 150) and human ELL3-rescued (n = 33) mouse oocytes. g, Schematic diagram of ELL3 TRIM-Away assay in mouse oocytes. h, Representative images showing ELL3 TRIM-Away efficiency, assayed by the validated ELL3-pAb, and and spindles and chromosomes in control and ELL3 TRIM-Away mouse oocytes. Scale bar, 20 μm. i, Percentage of abnormal MI spindles in control (n = 24) and ELL3 TRIM-Away (n = 31) mouse oocytes. j, Percentage of different types of spindle defects in ELL3 TRIM-Away (n = 31) mouse oocytes. Type I-short; type II-aMTOC clustering defects; type III-pole defocus; type IV-elongated; type V-chaos. k, Representative images of MII spindle morphologies in control and Ell3 KD mouse oocytes. Green, microtubules; blue, DNA. Scale bar, 20 μm. l, Percentage of abnormal MII spindles in control (n = 52) and Ell3 KD (n = 40) mouse oocytes. m, Representative images of ELL3-RFP localization at MI stage after cold-treatment. Red, RFP; green, microtubules; blue, DNA. Scale bar, 20 μm. Data are presented as mean ± s.e.m. (b,d,f,l). Data are from three independent experiments (b,d,f,i,l). N indicated the cumulative number of independent oocytes and independent experiments are denoted by dots (d,f,l). Statistical analysis was performed using a two-tailed, unpaired Student’s t-test (b,d,f,l) or a two-sided, fisher’s exact test (i). ns, not significant.
