qRT-PCR assay and RNA-seq

Testes samples were collected from WT and Neurog-cre Rbm46 KO at P10, and RNA was extracted using TRIzol reagent (Invitrogen). cDNA was prepared from 1 μg of total RNA through reverse transcription using a PrimeScriptRT Master Mix (TaKaRa). Diluted cDNA (1 μl) was used for each reaction using SYBR Green Premix Ex Taq II (RR820A, TaKaRa). A standard 20-μl reaction volume contained forward and reverse primers (200 nM), 1 μl of cDNA, and 10 μl of SYBR Green Mix. For RNA-seq, strand-specific libraries were prepared using the TruSeq Stranded Total RNA Sample Preparation Kit (Illumina) according to the manufacturer’s instructions before submitting to the Illumina NovaSeq 6000 system. The adaptor sequence and sequences with low quality were subsequently removed, and clean reads were then mapped to the mouse genome (mm10) with STAR (v 2.7.6A) with a GTF file downloaded from GENCODE database (57). Gene expression was evaluated using HTSeq, followed by DESeq2 normalization to evaluate gene expression. To define differentially expressed genes, the parameters (P < 0.05, fold change > 1.5) were used. RNA-seq data have been deposited in the Gene Expression Omnibus database ({"type":"entrez-geo","attrs":{"text":"GSE197282","term_id":"197282"}}GSE197282).