In situ hybridization of fancl in ovaries

The in situ hybridization on cryosections of presumptive ovaries were performed as previous described [56 (link)]. Sense and anti-sense digoxigenin-labeled cRNAs of fancl were synthesized and used in this study. The cDNA fragment of 786 nt containing the PHD domain of fancl was used to synthesize probe as previously described [38 (link)]. The in situ hybridization were photographed using a Nikon Eclipse Ni-U microscope (Nikon, Tokyo, Japan). Staining intensity in germ cells were quantified by analyzing the gray values using Image J software (National Institutes of Health, Bethesda, MD, USA). There were three independent replicates for the fish of each genotype.