GSM9251559: Tph cells, spleen,CD8 T cell-depleted PBMC-transferred, 2; Homo sapiens; RNA-Seq
The spleens were collected from immunodeficient mice transplanted with either human PBMCs depleted of CD8 T cells oe HSCs. Spleens were mechanically dissociated using gentle MACS dissociator (Miltenyi Biotec) and then lysed using HLB solution to remove red blood cells and obtain cell suspensions of mononuclear cells. Finally, the cells were resuspended in fluorescence-activated cell sorting (FACS) buffer and used cell sorting. RNA-sequencing was performed by isolated cells described in flow cytometry section. Total RNA was isolated from each sample using RNeasy Micro Kit (QIAGEN). The integrity and quantity of the total RNA were measured with an Agilent 2100 Bioanalyzer RNA 6000 Pico Kit (Agilent Technologies). Library preparation was performed using a NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolab) with NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (New England Biolabs) following the manufacturer's instructions. The quality of the libraries was assessed using an Agilent 2200 TapeStation High Sensitivity D1000 (Agilent Technologies).