GSM4697401: Atg7∆/∆  LP3; Mus musculus; RNA-Seq
Resected mouse tumors from Atg7+/+ and Atg7∆/∆ hosts were placed in complete media supplemented with 10% FBS and kept on ice.  Dissociation of the tumor tissue was carried out using a commercially-available mouse tumor dissociation kit and the Miltenyi GentleMACS dissocation system (Miltenyi Biotec, Cologne, Germany).  Isolated cells were filtered through a 40mm mesh strainer and PBMC's were isolated by density-dependent centrifugation.  Cells were washed complete media (10% FBS), then counted and assessed for viability using a Countess II cell counter (Invitrogen). Cells were then suspended to 1x10^6 per ml in complete media (10% FBS) for single cell emulsion preparation. scRNA-seq libraries were prepared according to 10X Genomics specifications (10X Genomics, Pleasanton, CA).  Independent cell suspensions were loaded for droplet-encapsulation by the Chromium Controller (10X Genomics). Single-cell cDNA synthesis, amplification, and sequencing libraries were generated using the Single cell 5' Reagent Kit following the manufacturer's instructions.  The libraries were shipped on dry ice to Novogene (Sacramento, CA), and sequenced in a single lane of an Illumina NovaSeq 6000 (Settings: Read1: 26, i7: 8, Read2: 91, for 200M bp paired-end reads per sample).