GSM8515122: lamina_propria_CD4_Tcells_rep2_mRNA; Mus musculus; RNA-Seq
Lamina propria CD4+ T cells (cell count of 2.5 × 10^4) were isolated from each sample using a FACS AriaII (BD) cell sorter. All samples were labeled with TotalSeqC hashtags antibodies (BioLegend), pooled, and encapsulated into droplets using a Chromium Single Cell Controller (10X Genomics), and libraries were prepared using Chromium Single Cell 5' Reagent Kits v2 (Dual Index) with Mouse TCR Amplification Kit and 5' Feature Barcode Kit (10X Genomics). The Controller was loaded with 2.5 × 10^4 cells per lane. mRNA, VDJ (TCR) and HTO libraries were prepared following the 10X Genomics user guide. scRNA-seq and TCR repertoire (VDJ region) sequencing of colonic lamina propria-specific CD4+ T cells. The scRNA-seq and TCR libraries were coupled with hashtag oligo (HTO) libraries for labeling the samples in order to demultiplex the pooled libraries into wild-type and Rag1 mutant mice at 3/12 weeks of age and with/without ANMV treatment cell populations for downstream analysis. The number of libraries sequenced is six for scRNA-seq, six for TCR (VDJ) repertoire, and six for HTOs.