title: GSM9443773: Foxp3YFP-CreKmt2d+/+, replicate 1, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Spleen CD4+FoxP3+ Treg cells from Foxp3YFP-CreKmt2d+/+ and Foxp3YFP-CreKmt2dfl/fl mice were sorted. The sorted Treg cells from three Foxp3YFP-CreKmt2d+/+ mice were pooled into one sample, while Treg cells from three Foxp3YFP-CreKmt2dfl/fl mice were combined into a separate sample for scRNA-seq. Library preparation and sequencing: The scRNA-seq libraries were prepared using the 10x Genomics Chromium Controller and Chromium Single Cell 3' V3.1 Reagent Kits (10x Genomics). Cells were concentrated to approximately 1,000 cells/uL and loaded into each channel to generate single-cell Gel Bead-In-Emulsions (GEMs). Following the reverse transcription step, GEMs were disrupted, and the barcoded cDNA was purified and amplified. The amplified cDNA was fragmented, A-tailed, ligated with adaptors and subjected to index PCR amplification. The final libraries were quantified using the Qubit High Sensitivity DNA assay (Thermo Fisher Scientific), and their size distribution was assessed using a High Sensitivity DNA chip on a Bioanalyzer 2200 (Agilent). All libraries were sequenced on an Illumina sequencer using a 150 bp paired-end run.