Illumina HiSeq 2000 paired end sequencing; Testis-to-ovary reprogramming and testis regression after ablation of Sox9 in Sox8-/- mice
Testis-to-ovary reprogramming and testis regression after ablation of Sox9 in Sox8-/- mice
Previously generated Sox9f/f; Sox8-/- mice were bred to Wt1-CreERT2 mice and the resulting double  heterozygous offspring harboring the Cre allele was backcrossed to Sox9f/f; Sox8-/- mice to  obtain heterozygous and homozygous compound Sox9; Sox8 conditional mutants. The same mating scheme was followed with the Sox9-CreERT2 mouse line. To  report CRE activity, an R26R-EYFP reporter allele was crossed into Wt1-CreERT2; Sox9f/f; Sox8-/- and Sox9-CreERT2; Sox9f/f; Sox8-/- mice.   Mice were feeded with a Tamoxifen (TX) supplemented diet (40mg TX/100g Harlan 2914 diet) for one month. TX administration were started at 2 months (P60). Both testes were extracted from six 5 months old, TX-treated mutant males (three Wt1- CreERT2; Sox9f/f; Sox8-/- and three Sox9-CreERT2; Sox9f/f; Sox8-/-). As controls, both gonads  were also extracted from four 5 months old control male mice (two TX-treated and two  untreated) as well as from two 4-5 months old normal females. All TX-treated mice were  euthanized three months after the initiation of diet TX-treatment for one month. The two  gonads of each individual were pooled, homogenized in 1ml of RNAzol (Molecular Research  Center, Inc.) per 100mg of tissue and the total RNAs were then individually purified from the  twelve samples following the RNAzol manufacturer’s instructions. After successfully passing  Macrogen Inc. quality check, the twelve RNAs were paired-end sequenced
Experimental Factor: Sox9+/+;Sox8-/-
genotype
Experimental Factor: Male
obsolete_sex