Targeted immune genes RNA-seq
tumors
TUMORS
Total RNA was extracted from 5 μm FFPE slides from pretreatment tumors using RNeasy FFPE Kit (Qiagen, catalog no. 73504) and quantified by Nanodrop and Qsep-100 (Thermo Fisher Scientific). The qualified RNA samples were hybridized with all the probes in the panel (NanoString Technologies). Finally, the hybridized products were purified through nCounter Prep Station to remove excess capture and reporter probes and to immobilize transcript-specific ternary complexes on a streptavidin-coated cartridge. Purified samples were finally scanned by nCounter Digital Analyzer (NanoString Technologies), analyzed used nSolver analysis software (v.4.0.70) and the nSolver Advanced data analysis package (v.2.0.134).