Transcriptome analysis of tumor infiltrating NK and T cells in CT26-bearing BALB/C mice

CT26 tumors were implanted subcutaneously into syngeneic BALB/C mice and allowed to grow for 15-25 days. Tumors were collected, mechanically dissociated, and immune cells enriched using Percoll gradient. Cells were then stained with viability dye, CD45, CD3, and NKp46 to FACS sort for T cells (Live/CD45+/CD3+/NKp46-) and NK cells (Live/CD45+/CD3-/NKp46+). Isolated tumor infiltrating NK and T cells were then processed for RNA sequencing analysis. Overall design: #1 - Pooled from N=4 mice, #2 - Pooled from N=6 mice

将CT26肿瘤皮下接种至同基因BALB/C小鼠，待肿瘤生长15~25天。收集肿瘤组织并进行机械解离，随后采用Percoll梯度（Percoll gradient）富集免疫细胞。使用细胞活性染料、CD45、CD3及NKp46对细胞进行染色，通过荧光激活细胞分选（Fluorescence-Activated Cell Sorting，简称FACS）分选出T细胞（活细胞/CD45+/CD3+/NKp46-）与NK细胞（活细胞/CD45+/CD3-/NKp46+）。将分离得到的肿瘤浸润NK细胞与T细胞进行RNA测序（RNA sequencing）分析。实验整体设计：样本#1由4只小鼠的组织混合制备，样本#2由6只小鼠的组织混合制备。