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The cyp17a1-/-;dmrt1-/- fish exhibited increased expression of fancl. (A and B) Relative expression of fancl in control fish, dmrt1-/- fish and cyp17a1-/-;dmrt1-/- fish at 17 dpf and 23 dpf was tested with qPCR. For fish RNA sampling at 17 dpf and 23 dpf, every 5 body trunks of fish collected were mixed into one sample, and 3 samples were examined. (C and D) In situ hybridization was performed on cryosections of presumptive ovaries from control fish and cyp17a1-/-;dmrt1-/- fish at 25 dpf using the antisense probe of fancl. Arrows point to the immature oocytes. (E) Comparison of differentially expressed gene in the ovaries of control fish and cyp17a1-/-;dmrt1-/- fish at 80 dpf. Volcano plot shows genes that were differentially expressed in ovaries between control and cyp17a1-/-;dmrt1-/- fish. (F) Gene set enrichment analysis based on genes differentiated expressed in cyp17a1-/-;dmrt1-/- fish at 80 dpf. The length of the bar represents the false discovery rate (FDR). (G) Relative expression of fancl in the ovaries of control fish and cyp17a1-/-;dmrt1-/- fish at 80 dpf with qPCR. For fish at 80 dpf, every three dissected ovaries were mixed as one sample, and 3 samples were examined. (H) The effect of Dmrt1 and DHT/Ar in regulating the relative luciferase activity driven by fancl promoter. DHT, dihydrotestosterone. ***, p < 0.001. Different letters in the bar chart represent significant differences.
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