title: Transcriptomic analysis of tumor-infiltrating NK cells from mifepristone-treated and vehicle-treated LLC-OVA tumor–bearing mice
design: Transcriptomic analysis of tumor-infiltrating NK cells from mifepristone-treated and vehicle-treated LLC-OVA tumor–bearing mice
library_name: Sample 5_p
library_construction_protocol: Tumors were excised from LLC-OVA tumor–bearing C57BL/6J mice on day 19. Each tumor was mechanically minced and enzymatically digested using IMDM supplemented with 10% FBS, collagenase D (1 mg/mL), collagenase A (1 mg/mL), and DNase I (0.4 mg/mL). Digestion was performed at 37 °C for 30–40 minutes with gentle agitation. Collagenase activity was terminated by adding EDTA (5 mM) to the samples. The resulting suspension was filtered through 70 µm cell strainers to obtain a single-cell suspension. Tumor-infiltrating NK cells were isolated by flow cytometric sorting based on CD3⁻ CD19⁻ CD56⁺ immunophenotype and collected in RNase-free tubes for downstream processing. Total RNA was extracted from sorted NK cells using the Qiagen RNA extraction kit according to the manufacturer's instructions, including optional on-column DNase digestion. RNA concentration and purity were determined using a NanoDrop spectrophotometer, and RNA integrity was assessed using Bioanalyzer. Only high-quality RNA samples were used for library construction. RNA-seq libraries were prepared by Novogene using their standard mRNA-seq library preparation pipeline.
identifiers.namespace: European Bioinformatics Institute
identifiers.id: E-MTAB-16381:Sample 5_p