Single-cell transcriptomic analysis of skin sorted CD45 positive and CD45 negative population from Il2ra upstream enhancer deleted (UP_Del) mice presenting with and without alopecia phenotype.

To understand the function of CD25/IL-2Ra in the regulation of alopecia development, we sorted live CD45 + and CD45- population from the skin of Up_del mice and were subjected to single-cell RNA-sequencing. Whole skins from UP_Del mice with and without alopecia phenotype along with C57BL/6 WT mice were harvested. Hypodermal layers were removed and remaining epidermal and dermal layer were subjected to mincing followed by mechanical force and enzymatic treatments. Cells were liberated after enzymatic digestion of the tissues. Cells were stained with CD45 antibody and live CD45+ and CD45- population were sorted by fluorescence-activated cell sorting (FACS). Their transcriptomics changes assessed by single-cell RNA-sequencing using the 10X genomics platform.