GSM6886467: 747495 WT-PEPD-mix_HTO-TSB; Mus musculus; RNA-Seq
All mice are on a C57BL/6 background. 10% WT CD45.1+ splenic cells were spiked into 1 WT CD45.2+ and 1 Pepd-/- CD45.2+ sample, and 10% WT CD45.1+CD45.2+ splenic cells were spiked into 1 WT CD45.2+ and 1 Pepd-/- CD45.2+ sample. Total T cells from the 4 samples described above and 1 WT CD45.2+ and 1 Pepd-/- CD45.2+ splenic samples were isolated by magnetic sorting, and then the 6 samples were multiplexed using Biolegend TotalSeq-B hashing antibodies by protocol (B0301 #155831, B0302 #155833, B0303 #155835, B0304 #155837, B0305 #155839, B0306 #155841). Pooled hashtagged cells were labelled with TotalSeq-A antibodies by protocol (anti-CD4 #100569, anti-CD8a #100773, anti-CD279 (PD-1) #109123, anti-CD44 #103063, anti-CD62L #104451, anti-CD25 #102055, anti-CD127 (IL7R), #135045, anti-ICOS #313555, anti-CD69 #104546, anti-CD27 #124235, anti-CD5 #100637, anti-KLRG1 #138431, anti-CD95 #152614, anti-CD45.1 #110753 and anti-CD45.2 #109853). After washing, cells were loaded into the chromium controller (10X-Genomics) chip and sequenced as previously described (16). Reads for TotalSeq anti-CD45.1 and anti-CD45.2 alone could not be detected, thus the CD45.1+ and CD45.1+/CD45+ populations could not be separated from the CD45.2 in this scRNAseq experiment dataset. Cells were loaded into the chromium controller (10×-Genomics) chip following the standard protocol for the chromium single cell 3' kit. (Multi-omic single cell analysis resolves novel stromal cell populations in healthy and diseased human tendon., Kendel et al. Scientific Reports., 2020) Total-Seq-A ADT and Total-Seq-B Hashtag-seq as previously described following biolegend protocols (Multi-omic single cell analysis resolves novel stromal cell populations in healthy and diseased human tendon., Kendel et al. Scientific Reports., 2020)