Gestational estrogenic and anti-androgenic endocrine disruption alters stromal-epithelial remodeling during mammary gland involution in rats.

RNA was extracted from 100 mg of inguinal mammary gland tissue from three randomly selected samples per group, following the manufacturer’s instructions from the Aurum Total RNA Fatty and Fibrous Tissue Kit (Bio-Rad, Ontario, Canada, Cat. 7326830) 1,2,4. RNA quality was assessed by Nanodrop (Thermo Scientific), and Agilent 2100 Bioanalyzer’s RNA 6000 Nano Kit was used to confirm and validate RNA integrity and concentration (all samples were RIN > 8). The extracted RNA samples were sent to the Genomic Centre of the CERMO-FC – Université du Québec à Montréal (UQAM) to undergo RNA sequencing on a Illumina NovaSeq PE100 (3200M reads). For sequencing data analyses, raw sequencing reads underwent a quality assessment with FastQC v0.12.0, trimming with Trimmomatic v0.39, and aggregating with MultiQC v1.20. High-quality trimmed reads were aligned to the Rattus norvegicus Rnor_6.0 reference transcriptome (https://may2021.archive.ensembl.org/Rattus_norvegicus) with HISTA2 v2.2.1, the alignment processing were used SAMtools v1.22.1, for transcript assembly and quantification were carried out with StringTie v2.1.5.