GSM8031539: mammary_DN_NK11_Tcells_rep1; Mus musculus; RNA-Seq
scRNA-Seq: Mammary lymphoid cells (CD45+CD90.2+) from pregnant (G18.5) and lactating (L5) mice were sorted from samples labeled with TotalSeqC hashtag oligos using a Sony MA900 sorter. Likewise, the mammary epithelial cells and myoepithelial cells from samples labeled with TotalSeqC hashtags oligos (Biolegend) with a Sony MA900 sorter. Sorted cells were pooled together and loaded to a Chromium Single Cell Controller (10X Genomics) to encapsulate the cells into droplets. Bulk RNA-Seq: T cells were sorted with a Sony MA900 sorter from the mammary glands of pregnant (E18.5) mice. The RNA from these sorted T cells were extracted using RNeasy Micro Kit (Qiagen) according to manufacturer's instructions and libraries were prepared using Tecan's Solo mouse kit according to manufacturer's instructions. scRNA-seq libraries were prepared using 10X scRNAseq 5' v2 with hashtag oligos (HTOs) and VDJ (T cell receptor) sequences, whereas the bulk RNA-Seq libraries prepared using Tecan's Solo mouse kit. All libraries were prepared according to the manufacturer's instructions. scRNA-seq on mammary epithelial-myoepithelial and lymphoid cells in mice. Bulk RNA-Seq on CD8 beta+ T-cells and DN (double negative) NK1.1+ lymphocytes in mice. The scRNA-seq libraries were coupled with hashtag oligo (HTO) libraries for labeling the samples in order to demultiplex the pooled RNA libraries into individual mice for downstream analysis, whereas the lymphoid cells were also used for generating T cell receptor (VDJ region) sequencing libraries for characterizing the immune repertoire landscape.