GSM5575729: AB12898; Mus musculus; RNA-Seq
Isolation of tumor infiltrating leukocytes:  Tumor bearing mice were sacrificed 7 days days after treatment injection inoculation. The tumors underwent mechanical (gentleMACSTM C tube, Miltenyi Biotec Inc., San Diego, CA) and enzymatic digestion (0.1mg/ml DNase type I (Roche), and 1mg/ml Collagenase IV (Worthington) in RPMI-1640) for 15 min at 37°C. Cells then filtered through 100µm cell strainer, washed with ice cold sorting buffer, centrifuged (5 min, 4°C, 300g), and stained with fluorophores conjugated antibodies. LN and spleen were harvested 3 hours after treatment injection.Lymph nodes and spleen were dissected through a 70 µm nylon cell strainer and washed with PBS. Samples were flow sorted using AriaIII (BD Biosciences, BD Diva Software) cell sorter. 3' end mRNA libraries were prepared for sequencing using MARS-seq (Jaitin et al, Science 2014)