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Incubations of the cells were carried out in microplates at a density of 1.5 10 4 cells per well in HBSS buffer (Invitrogen) with 20 mM HEPES (pH 7.4) and 500 M IBMX

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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)
