GSM9530363: control; Mus musculus; RNA-Seq
Recombinant mouse IL-17A was administered to OCCC-bearing mice, and ovarian tumors were harvested 7 days after treatment. Tumor tissues were finely minced with scissors and enzymatically digested with Collagenase D (1.25 mg/mL; Merck, Darmstadt, Germany) at 37°C for 30 minutes. The resulting single-cell suspensions were stained with an anti-CD45 antibody (clone 30-F11, 1:800; BioLegend, San Diego, CA, USA) and 7-aminoactinomycin D (7-AAD, 1:200; BioLegend), and viable CD45⁺7-AAD⁻ cells were sorted using a BD FACSAria II cell sorter (BD Biosciences, San Jose, CA, USA). Sorted cells were loaded onto a BD Rhapsody Cartridge and processed using the BD Rhapsody Cartridge Reagent Kit (BD Biosciences) according to the manufacturer's instructions. Single-cell cDNA synthesis was performed by Immunogeneteqs (Chiba, Japan) using the BD Rhapsody cDNA Kit (BD Biosciences). Whole Transcriptome Analysis (WTA) libraries were generated using the Immune Response Panel Mm and sequenced on a NovaSeq 6000 S4 flow cell (NovaSeq 6000 S4 Kit v1.5; Illumina, San Diego, CA, USA), with a sequencing depth of approximately 20,000 reads per cell. Sorted cells were loaded onto a BD Rhapsody Cartridge and processed using the BD Rhapsody Cartridge Reagent Kit (BD Biosciences) according to the manufacturer's instructions. Single-cell cDNA synthesis was performed by Immunogeneteqs (Japan) using the BD Rhapsody cDNA Kit (BD Biosciences). Whole Transcriptome Analysis (WTA) libraries were generated using the Immune Response Panel Mm and sequenced on a NovaSeq 6000 S4 flow cell (NovaSeq 6000 S4 Kit v1.5; Illumina).