Milrinone

A rabbit polyclonal anti-KIF2A antibody was purchased from Novus Biologicals (Littleton, CO, USA). Normal rabbit IgG was purchased from Santa Cruz Biotechnology (Dallas, CA, USA). A rabbit polyclonal anti-SKA1 antibody was purchased from Abcam (Cambridge, MA, USA). A rabbit monoclonal anti-TACC3 antibody was purchased from Abcam (Cambridge, MA, USA). Mouse monoclonal FITC-conjugated anti-α-tubulin antibody, TRITC-conjugated phalloidin, nocodazole and taxol were purchased from Sigma-Aldrich (St Louis, MO, USA). Sheep polyclonal anti-BubR1 antibody was purchased from Abcam (Cambridge, MA, USA). Alexa Fluor 546-conjugated donkey anti-sheep and donkey anti-rabbit antibodies were obtained from Invitrogen (Carlsbad, CA, USA). M2 medium was purchased from Sigma-Aldrich (St Louis, MO, USA). Milrinone was obtained from Cayman Chemical Co. (Ann Arbor, MI, USA). Mounting medium containing DAPI was obtained from Vector Laboratories (Burlingame, CA, USA). Horseradish peroxidase (HRP)-conjugated mouse anti-rabbit IgG and horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG were obtained from Zhong Shan Jin Qiao Co. (Beijing, P. R. China). A mouse monoclonal anti-β-actin antibody was obtained from Abcam (Cambridge, MA, USA). Enhanced chemiluminescence (ECL) detection substrate was obtained from Xiamen Lulong Biotech Co. (Xiamen, P. R. China).

A rabbit polyclonal anti-Axin-1 antibody was purchased from Millipore (Billerica, MA, USA). Sheep polyclonal anti-BubR1 and rabbit polyclonal anti-Nek9 antibodies were from Abcam (Boston, USA). Mouse monoclonal anti-γ-tubulin antibody, anti-α-tubulin FITC-tagged antibody, and TRITC-tagged anti-phalloidin were obtained from Sigma-Aldrich (St Louis, USA). Alexa Fluor 546-conjugated donkey anti-rabbit and donkey anti-sheep antibodies were purchased from Life Technologies (Carlsbad, USA). An Alexa Fluor 546-conjugated donkey anti-mouse antibody was purchased from Invitrogen Life Technologies (Carlsbad, USA). Control rabbit IgG was purchased from Santa Cruz Biochemicals (Dallas, USA). Mounting medium containing DAPI was purchased from Vector Laboratories (Burlingame, USA) and milrinone was purchased from Cayman Chemical Co. (Ann Arbor, USA).

Mice were given 10 IU pregnant mare serum gonadotropin (PMSG; Ningbo Sansheng Pharmaceutical Company, China) intraperitoneally. After 48 h, the mice were killed and ovaries were removed. GV-intact oocytes were collected in M2 medium (Sigma, USA) and were cultured under paraffin oil in a 5% CO2, 95% air incubator. Oocytes to be blocked at the GV stage were cultured in M2 medium plus 2.5 mM milrinone (Cayman, USA), whereas those allowed to mature in vitro were cultured in fresh M2 medium for 8.5 or 12.5 hr. For parthenogenesis, in vitro matured MII oocytes were activated with 10 μM SrCl2 for 4–6 hours. The oocytes were then observed with light microscopy to observe the pronucleus; the activated eggs were then cultured in the KSOM medium to the 2-cell embryo stage.