Digoxigenin–uridine triphosphate–labeled and fluorescein-labeled antisense RNA probes were transcribed using the MEGAscript Kit (Ambion) according to the manufacturer’s instructions. WISH was performed according to previously published methods (39). For double in situ hybridization, anti–digoxigenin-peroxidase (POD) (11633716001, Roche) and anti–fluorescein-POD (11426346910, Roche) were used as primary antibodies to detect digoxigenin-labeled pinhead probe and fluorescein-labeled gsc probe, respectively.

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