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. 2022 Dec 7;22(1):100481. doi: 10.1016/j.mcpro.2022.100481

Fig. 3.

Fig. 3

CEP57L1 Regulates the Assembly of Meiotic Apparatus in Mouse Oocytes.A, gene ontology analysis of the differentially expressed proteins with enrichment in the regulation of cell cycle. The four complexes are marked with yellow circles. The red circle indicates CEP57L1. B, relative abundance of CEP57L1 in oocytes. C and D, relative fluorescence intensity of CEP57L1 in GV oocytes and MII oocytes. E, cellular distribution of CEP57L1 in oocytes at GV, GVBD, and MI stages. Arrowheads point to CEP57L1 signals. The scale bar represents 20 μm. F, schematic presentation of the experimental design to investigate the role of CEP57L1 in meiosis. G, knockdown efficiency of two Cep57l1 siRNAs. H, representative images of oocytes from control and two siCep57l1 groups. Arrowheads point to oocytes that fail to extrude a polar body or oocytes with the abnormal polar body. The scale bar represents 50 μm. I, rate of Pb1 extrusion in control oocytes and two siCep57l1 groups. J, representative images of the spindle and chromosome morphology of MI oocytes stained with α-tubulin antibody (green) and with propidium iodide (red), respectively. K, quantification of control and two siCep57l1 groups with aberrant spindles. The scale bar represents 20 μm. L, representative images of kinetochore-microtubule (K–M) attachment in control and two siCep57l1 groups. Oocytes were costained with a-tubulin-FITC antibody (microtubules, green), CREST antibody (kinetochore, purple), and Hoechst 33342 (chromosomes, blue). M, quantitative analysis of the defective K–M attachment in the control group and two siCep57l1 groups. N, representative chromosome spread images of oocytes stained with BubR1 antibody (green) and propidium iodide (red) in the control group and two siCep57l1 groups. O, quantitative analysis of the BubR1 intensity in the control group and two siCep57l1 groups. Data are expressed as the mean ± SD from three independent experiments in which at least 100 oocytes were analyzed for each group. For statistical analysis, a two-tailed Student’s t test was used in all panels, compared with GV or control.