RNA-seq of TC-1-A9 tumors, 9 days after IT
Cell lines or excised tumors were immediately placed into the RNAlater Solution (Thermo Fisher Scientific, Waltham, MA, USA) and stored at 4C for a maximum of 14 days. Samples were disrupted by a rotor-stator homogenizer (Omni TH, Kennesaw, GA, USA) in a lysis buffer and total RNA was isolated using the NucleoSpin RNA kit (Macherey Nagel, Dren, Germany). NGS libraries were prepared using the QuantSeq 3mRNA-Seq Library Prep Kit (FWD) (Lexogen, Vienna, Austria). Sequencing was performed on the NextSeq 500 System (Illumina, San Diego, CA, USA) yielding unpaired 75-bp reads.