Transcriptome profiling of FGL2-stimulated enteric neural crest cells to investigate metabolic dysregulation in Hirschsprung disease pathogenesis

This dataset comprises bulk RNA sequencing data from primary mouse enteric neural crest cells (ENCCs) stimulated with fibrinogen-like protein 2 (FGL2) or left untreated as controls. ENCCs were isolated from embryonic day 14–15 C57BL/6J mice and cultured under defined conditions to ensure >95% expression of P75 and nestin markers. Following FGL2 stimulation, total RNA was extracted, and sequencing libraries were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina. Paired-end sequencing (2 × 150 bp) was performed on the Illumina NovaSeq 6000 platform. The study aims to identify transcriptional changes and signaling pathway alterations induced by FGL2 in ENCCs, providing insights into the role of FGL2-mediated metabolic dysregulation in the pathogenesis of Hirschsprung disease. Overall design: Primary ENCCs were obtained from gut tissues of C57BL/6J mouse embryos at gestational day 14–15. Cells were cultured in complete mouse ENCC medium until reaching 80–90% confluence and confirmed to express P75 and nestin in >95% of cells by immunofluorescence. Cells were randomly assigned to two groups: FGL2-treated group – ENCCs stimulated with recombinant FGL2 protein. Control group – ENCCs cultured under identical conditions without FGL2. Three biological replicates were included per group. Total RNA was extracted using TRIzol reagent, and RNA quality was assessed on an Agilent 2100 Bioanalyzer. Poly(A)+ mRNA was enriched, fragmented, and reverse-transcribed to cDNA. Libraries were constructed using the NEBNext Ultra RNA Library Prep Kit and sequenced (paired-end, 2 × 150 bp) on an Illumina NovaSeq 6000.