GSM9398977: LV-1A10-2; Homo sapiens; RNA-Seq
Total RNA was extracted from 5×10⁶ cells collected from each group (MOCK-T, LV-CAR-T, and CISH-KO-CAR-T) using Trizol reagent (Invitrogen) following the manufacturer's instructions. RNA integrity and quality were verified by agarose gel electrophoresis and assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies). For library preparation, eukaryotic mRNA was first enriched with Oligo(dT) beads, then fragmented and reverse-transcribed into cDNA using the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB #7530). The resulting double-stranded cDNA fragments underwent end repair, adenylation, and Illumina adapter ligation. After purification with AMPure XP Beads (1.0X) and PCR amplification, the constructed libraries were subjected to paired-end sequencing on an Illumina Novaseq 6000 platform at Gene Denovo Biotechnology Co. (Guangzhou, China).