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. 2021 Sep 17;9:749116. doi: 10.3389/fcell.2021.749116

FIGURE 2.

FIGURE 2

TRF1 depletion impairs kMT attachment and increases the rate of aneuploidy. Oocytes injected with dsEGFP or dsTRF1 were cultured in medium containing IBMX for 24 h and then transferred to IBMX-free medium for 8 h. (A–D) Oocytes at the MI stage were fixed and stained with anti-α-tubulin antibody and DAPI (scale bar, 10 μm). Spindle abnormality, chromosome misalignment, and metaphase plate width were quantified and are shown in representative images. Data are presented as mean ± SEM from three independent experiments. (E–G) After cold treatment, oocytes were fixed and stained with anti-centromere antibody (ACA), anti-α-tubulin antibody, and DAPI to visualize kinetochore, spindle, and DNA, respectively. The percentages of unattached kinetochores per oocyte and oocytes with abnormal kMT attachment were quantified, also shown in representative images (scale bar, 10 μm). Data are presented as mean ± SEM from three independent experiments. (H,I) Chromosome spreading of TRF1-depleted MII oocytes. Kinetochores and DNA were stained with ACA and DAPI, respectively (scale bar, 20 μm). The incidence of aneuploidy was quantified. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001.