Transcriptomic profiling of TGF-ß–stimulated primary mouse cardiac fibroblasts
Primary cardiac fibroblasts were isolated from neonatal C57BL/6J mouse hearts and stimulated with TGF-ß1 in vitro to induce a profibrotic phenotype. Bulk RNA sequencing was performed on fibroblasts treated with TGF-ß for 48 hours and on matched untreated controls to characterize transcriptional programs associated with TGF-ß–driven fibroblast activation. Differential expression and gene ontology analyses were used to identify pathways related to extracellular matrix remodeling, TGF-ß signaling, and DNA damage responses. These data support mechanistic studies on CRABP2–MRE11–MRN–dependent regulation of cardiac fibroblast activation and fibrosis. Overall design: Primary cardiac fibroblasts were obtained from 1-day-old C57BL/6J mice and cultured under standard conditions. Cells were divided into 2 groups: (1) untreated controls and (2) cells treated with recombinant TGF-ß1 (10 ng/mL) for 48 hours. For each condition, independent biological replicates of fibroblasts isolated from different litters were processed. Total RNA was extracted using TRIzol, and sequencing libraries were prepared using a standard poly(A)-enriched mRNA protocol and sequenced on an Illumina platform. Raw reads were aligned to the mouse reference genome (mm39), and differential gene expression was assessed between TGF-ß1–treated and control fibroblasts.