Single cell RNA-seq analysis of novel splenic DC

To understand the cellular diversity within CD11b+ splenocytes between control and GM-CSF-treated mice, we performed the CITE-seq in control and GM-CSF treated mice. Overall design: FACS sorted CD11b+ splenocytes were loaded into the Chromium system (10x Genomics, Pleasanton, CA, USA) targeting 7,000 cells per sample. The cDNA libraries for mRNA were generated using Chromium Single Cell 3' v3 Reagent Kits according to the manufacturer's instruction. Following the CITE-seq protocol, ADT PCR additive primers were added to cDNA PCR and the ADT libraries were generated separately from the mRNAs.为解析对照组与GM-CSF处理组小鼠CD11b阳性（CD11b+）脾细胞内的细胞异质性，我们对两组小鼠开展了转录组与表位联合测序（CITE-seq）。 实验设计概况：通过荧光激活细胞分选（Fluorescence-Activated Cell Sorting, FACS）富集的CD11b+脾细胞被上样至Chromium系统（10x Genomics，美国加利福尼亚州普莱森顿）中，每样本预设捕获目标细胞数为7000个。依照制造商说明书，使用Chromium单细胞3'端v3试剂试剂盒构建mRNA的cDNA文库。遵循转录组与表位联合测序实验流程，向cDNA PCR体系中加入抗体衍生标签（Antibody-Derived Tag, ADT）PCR附加引物，并独立于mRNA构建ADT文库。