title: GSM6705939: B16-OVA, Isotype IgG + TBK1i, Replicate 3; Mus musculus; RNA-Seq
library_construction_protocol: Tumors were dissected on day 14 post-inoculation and disassociated using the Miltenyi Mouse Tumor Dissociation Kit and gentleMACS Octo-Dissociator (Miltenyi) using the m-TKD-1 program. After filtering through a 70-μm filter, live cells were isolated using a density gradient with Lympholyte-M separation media (Cedarlane Labs) per the manufacturer's specifications. CD45+ tumor-infiltrating immune cells were enriched by positive selection with MicroBeads (Miltenyi) and magnetic separator (Miltenyi). Four representative samples from each of the Control (Vehicle/IgG-treated), aPD-1 (Vehicle/anti-PD-1-treated), TBK1i (TBK1i/IgG-treated), and αPD-1 + TBK1i (TBK1i/anti-PD-1) groups were selected and droplet-based isolation of single cells was performed with the Chromium Controller (10X Genomics) according to the manufacturer's specifications. Subsequent generation of 3′ sequencing libraries was performed per manufacturer's instructions (10X Genomics). Libraries were prepared utilizing Chromium Next GEM Single Cell Reagent Kits 3′v2 chemistry (10X Genomics). Characterization of the sequencing library was performed with TapeStation (Agilent) and Qubit (ThermoFisher) instruments. 