RNAseq of different CD176-TRUCKs before and after co-culture with A549 tumor cells
RNAseq of different CD176-TRUCKs before and after co-culture with A549 tumor cells
CD8+ untransduced T cells, CD176-iEGFP-, CD176-iIL18-, or CD176-iIL12-TRUCKs from 4 different donors, before or after 48 h co-cultures with A549 cells at a 1:1 ratio, and A549 cells cultured alone for 48 h were used for bulk RNA analysis. Suspension cells from each sample were resuspended Total RNA was isolated using RNeasy Plus Kit (QIAGEN, Hilden, Germany) according to manufacturer's instructions. The RNA sequencing library was generated from 300 ng total RNA using the NEBNext® Poly(A) mRNA Magnetic Isolation Module, followed by the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina (both from New England Biolabs®, Ipswich, MA, USA) according to the manufacturer's protocols, except that all reactions were downscaled to 2/3 of initial volumes.