The following antibodies and dilution were used for staining: rabbit anti-GFP (Medical & Biological Laboratories Co., Ltd, #598, 500 × ), anti-beta-galactosidase (MP Biomedicals, #0855976, 1000 × ), rat anti-DE-cad (DCAD2, 20 × )61 (link), mouse anti-Gasp (2A12, DSHB, 2 × ), anti-Serp (300 × ) and Verm (300 × )34 (link), anti-Pio (50 × )36 (link), anti-Arl3 (500 × )19 (link) and CBP-Alexa 488 (50 × )35 (link). Embryos were dechorionated with 1/2 diluted commercial bleach and were fixed by mixing vigorously in the mixture of 2 ml 4% paraformaldehyde in PBS and 2 ml heptane in liquid scintillation vial for 30 min at room temperature. After removal of heptane, embryos were devitelinized by addition of 2 ml methanol and vigorous mixing. Devitellinized embryos were washed two times with methanol and rehydrated with PBS. Embryos were stained as described previously56 (link). Blocking, antibody incubation and washing were performed with 1% BSA and 0.2% Triton X-100 in PBS. The anti-Katanin 60 antibody (100 × ) was generously donated by Naoyuki Fuse and Fumio Matsuzaki.
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