Ovarian Stroma RNA-seq of Progesterone receptor null vs WT mice at 8hrs post hCG

Progesterone receptor-dependent expression of the ovairan stroma is investigated to determine PGR control of stromal functions at ovulation. Ovaries were collected from 21-25 day old PGR(-/-) and PGR(+/+) littermates at 8hrs post-hCG. N=3-4 per genotype with each experimental replicate consisting of tissue pooled from 3 mice. The Stromal tissue of the ovary (GC-depleted ovary) was used for mRNA-seq Illumina Truuseg Strandard mRNA kit, Illumina Novaseq 600 SP. Paired end, 100bp, depth of 54-72M reads. Quality control checked using FastQC. Salmon lignment to GRCm39 M26 mouse transcriptome. DESeq2 used to asses differentially expressed genes.本研究旨在探究孕酮受体（Progesterone receptor, PGR）依赖的卵巢基质表达特征，以明确PGR对排卵时基质功能的调控作用。实验于人绒毛膜促性腺激素（human chorionic gonadotropin, hCG）处理后8小时，从21~25日龄的PGR敲除（PGR(-/-)）与野生型（PGR(+/+)）同窝仔鼠体内采集卵巢。每组基因型设置3~4个生物学重复，每个实验重复样本由3只小鼠的组织混合制备。提取卵巢基质组织（即去除颗粒细胞（granulosa cell, GC）的卵巢）进行mRNA测序：采用Illumina TruSeq Standard mRNA建库试剂盒，依托Illumina NovaSeq 6000 SP测序平台，实施双端100bp测序，测序深度为5400万至7200万条reads。使用FastQC进行测序数据质量控制，通过Salmon软件将测序数据比对至GRCm39 M26版本小鼠转录组，并利用DESeq2分析差异表达基因。