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The gonads were then incubated with anti-digoxigenin antibody (Sigma, #11333089001) O/N at 4C, followed by three washes in PBST, then developed in AP substrate BCIP/NBT (Sigma, #B5655)

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Editorial assistance was provided by Laura Brogelli, PhD, and Aashni Shah, BSc (Polistudium SRL, Milan, Italy)