Fig 2: Effects of Pds5A depletion on the K-M attachment and chromosome ploidy in mouse oocytes.(A) Fluorescence images kinetochores and microtubule fibers in control, Pds5A-MO, and Pds5A-rescue oocytes at MI stage. White arrows indicate unattached kinetochores. Scale bars, 5 and 2.5 μm. (B) The percentage of unattached kinetochores in control (n = 90), Pds5A-MO, (n = 125), and Pds5A-rescue (n = 103) oocytes at MI stage. (C) Representative images of chromosome spreads in control, Pds5A-MO, and Pds5A-rescue oocytes at MII stage. Scale bar, 8 μm. (D) The percentage of aneuploid oocytes at MII stage in control (n = 76), Pds5A-MO (n = 78), and Pds5A-rescue (n = 76) groups. Data in (B) and (D) were designated as mean percentage (means ± SEM) of at least three independent experiments. **P < 0.01; ***P < 0.001; ****P < 0.0001.

Fig 3: Effects of Pds5A depletion on the mouse oocyte meiotic progression.(A) Representative images of GVBD oocytes observed at 4 hours following release from 3-isobutyl-1-metyl-xanthine in control and Pds5A-MO groups. Scale bar, 80 μm. (B) The rate of GVBD in control (n = 169) and Pds5A-MO (n = 185) oocytes. (C) Representative images of MII oocytes observed at 10 hours post-GVBD in control, Pds5A-MO, and Pds5A-rescue (Pds5A-MO + Pds5A-cRNA) groups. Scale bar, 80 μm. (D) The rate of PBE in control (n = 119), Pds5A-MO (n = 119), and Pds5A-rescue (n = 113) oocytes. (E) Representative images of chromosome morphology in Pds5A-MO oocytes without PB1 after maturation. Scale bar, 5 μm. (F) The percentage of oocytes arresting at different developmental stages in Pds5A-MO (n = 76) group. (G) Fluorescence images of BubR1 present on the chromosomes in control and Pds5A-MO oocytes at Pro-MI and MI stages. Scale bar, 5 μm. (H) The proportion of BubR1 presence on the chromosomes in control (n = 73) and Pds5A-MO (n = 69) oocytes at MI stage. (I) Fluorescence images of Mad2 present on the chromosomes in control and Pds5A-MO oocytes at Pro-MI and MI stages. Scale bar, 5 μm. (J) The proportion of Mad2 presence on the chromosomes in control (n = 71) and Pds5A-MO (n = 86) oocytes at MI stage. Data in (B), (D), (F), (H), and (J) were designated as mean percentage (means ± SEM) of at least three independent experiments. **P < 0.01; ns, no significance.

Fig 4: The oocyte maturation and female fertility in Pds5A+/− mice.(A) Representative images of pups from WT and Pds5A+/− female mice. Scale bar, 1 cm. (B) The average number of pups per litter from WT (n = 9) and Pds5A+/− (n = 8) female mice. (C) Representative images of superovulated oocytes from WT and Pds5A+/− female mice. Scale bar, 80 μm. (D to F) The number, PBE rate, and death rate of superovulated oocytes in WT (n = 248) and Pds5A+/− (n = 173) groups. (G) Fluorescence images of spindles and chromosomes in WT and Pds5A+/− superovulated oocytes. Scale bars, 20 and 10 μm. (H) The rate of abnormal spindles in WT (n = 85) and Pds5A+/− (n = 84) superovulated oocytes. (I) The number of different defective spindles in Pds5A+/− (n = 40) superovulated oocytes. (J) The rate of misaligned chromosomes in WT (n = 85) and Pds5A+/− (n = 84) superovulated oocytes. (K) Representative images of chromosome spreads in WT and Pds5A+/− superovulated oocytes. Scale bar, 10 μm. (L) The percentage of aneuploid oocytes in WT (n = 84) and Pds5A+/− (n = 57) groups. (M) Fluorescence images of spindles and chromosomes in WT and Pds5A+/− MI oocytes. Scale bar, 20 μm. (N) The rate of abnormal spindles in WT (n = 100) and Pds5A+/− (n = 118) MI oocytes. (O) The number of different defective spindles in Pds5A+/− (n = 36) MI oocytes. (P) The rate of misaligned bivalents in WT (n = 100) and Pds5A+/− (n = 118) MI oocytes. Data in (B) and (D) were designated as means ± SD, and (E), (F), (H), (J), (L), (N), and (P) were designated as means ± SEM of at least three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Fig 5: Identification of binding partners of Pds5A.(A) Representative binding candidates of Pds5A as shown in MS data. (B) Costaining of Pds5A and Kif5B in oocytes at MI stage. Scale bars, 20 and 5 μm. (C) Fluorescence intensity profiles of Pds5A and Kif5B along the white line. (D) Co-IP of Pds5A and Kif5B as precipitated with Pds5A antibody. The blots of precipitates were probed with Kif5B and Pds5A antibodies, respectively. (E) Co-IP of Pds5A and Kif5B as precipitated with Kif5B antibody. The blots of precipitates were probed with Pds5A and Kif5B antibodies, respectively.