Rabbit anti tpx2

Oocytes were fixed in 3.8% paraformaldehyde containing 0.1% Triton X-100 (Sigma-Aldrich, St. Louis, MO, USA) for 1 h at 37 °C, and washed 3 times in blocking solution with 1x PBS containing 0.3% BSA and 0.01% Tween-20. Oocytes were incubated overnight with rabbit anti-α-tubulin (Cell Signaling Technology, Danvers, MA, USA), rabbit anti-TPX2 (Novus, Littleton, CO, USA), and rabbit anti-DAZL (Abcam, Cambridge, MA, USA) in blocking solution. Then, oocytes were washed 3 times with blocking solution and incubated with anti-rabbit IgG for 1 h at RT, mounted using Vectashield containing DAPI (Vector Laboratories, Burlingame, CA, USA), and imaged by confocal (Leica Microsystem, Buffalo Grove, IL, USA).

Cell lysis and WB were performed as described in reference 62 (link). Antibodies were as follows: rabbit anti-TPX2 (1:1,000; Novus Biologicals), mouse anti-Aurora-A (0.5 µg/ml; BD Transduction Laboratories), rabbit anti-pAurora-A (Thr288) (3079 clone C39D8, 1:100; Cell Signaling Technology, Inc.), and mouse anti-GAPDH (SC-32233, 1:1,000; Santa Cruz Biotechnology). HRP-conjugated secondary antibodies (Bio-Rad Laboratories S.r.l.) were revealed using the Clarity Western ECL Substrate (Bio-Rad Laboratories S.r.l.).