GSM8395216: MousePlaque_LL_female, mRNA; Mus musculus; RNA-Seq
Digested tissue was filtered through a 70-µm strainer, washed with cold PBS, centrifuged at 500 g for 5 min. at 4 °C and resuspended in PBS supplemented with 1% vol/vol BSA. Single-cell suspensions were placed on ice and stained with eFluor 780 Fixable Viability Dye (eBioscience, 65-0865-14) for 30 min., washed, blocked with TruStain FcX (anti-mouse CD16/32) Antibody (Biolegend, 101319) for 5 min. and then stained with PerCP/Cyanine5.5 anti-mouse CD45 Antibody (Biolegend, 103132) for 30 min. Live, CD45+ cells were purified by fluorescence-activated cell sorting using a BD FACSAria II instrument at the Cytometry and Cell Sorting Laboratory at the NYU Grossman School of Medicine at NYU Langone Health. For sorting, specimens were pooled by experimental group. The sorter was equipped with a 100 µm nozzle. scRNA-seq libraries were prepared using the Chromium Single Cell 3' Library v3 reagent, Gel Bead and Multiplex Kit and Chip Kit (10X Genomics; 1000268, 1000215 and 100127, respectively) according to the manufacturer's instructions.