title: GSM9162425: enteric neural crest cells sample9 treated with FGL2 recombination protein; Mus musculus; RNA-Seq
library_construction_protocol: 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. Total RNA was extracted using TRIzol reagent kit (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's protocol. RNA quality and integrity were assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) and verified by RNase-free agarose gel electrophoresis. Poly(A)+ mRNA was enriched from total RNA using Oligo(dT) beads. The enriched mRNA was fragmented into short fragments using fragmentation buffer and reverse-transcribed into cDNA using NEBNext Ultra RNA Library Prep Kit for Illumina (NEB #7530, New England Biolabs, Ipswich, MA, USA). The purified double-stranded cDNA fragments were end-repaired, A-tailed, and ligated to Illumina sequencing adapters. Adapter-ligated fragments were purified with AMPure XP Beads (1.0×) and PCR-amplified. The final cDNA libraries were sequenced on an Illumina NovaSeq 6000 platform (paired-end, 2 × 150 bp) by Gene Denovo Biotechnology Co., Ltd. (Guangzhou, China).