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

FIGURE 5.

FIGURE 5

Inhibition of inner centromere or kinetochore function decreases TRF1 level at telomeres. (A–C) Ndc80/Hec1 chromosome spreads were prepared from MI oocytes depleted Hec1 using the Trim-away method, and chromosome spreads were stained with anti-TRF1 antibody. Kinetochores and DNA were labeled with anti-centromere antibody (ACA) and DAPI, respectively (scale bar, 20 μm). (A) Representative images from three independent experiments are shown. (B,C) The normalized intensity of TRF1 at p-arms and q-arms was quantified. (D–F) Chromosome spreads were prepared from MI oocytes after treatment with DMSO (Control), AZ3146, or ZM447439 and stained with anti-TRF1 antibody. Kinetochores and DNA were labeled with anti-centromere antibody (ACA) and DAPI, respectively (scale bar, 20 μm). (D) Representative images from three independent experiments are shown. (E,F) The normalized intensity of TRF1 at p-arms and q-arms was quantified. (G,H) After treating with DMSO (Control), AZ3146, or ZM447439, oocytes were cultured for 12 h and subjected to telomere Q-FISH. Relative telomere FISH intensity was quantified and shown with representative images (scale bar, 20 μm). ns; not significant, ***p < 0.001.