title: GSM8974420: D667 GEX library 1; Homo sapiens; OTHER
library_construction_protocol: UpCell plates were incubated on ice for 20 min for non-enzymatic cell dissociation. Samples were stained for 30 min with TotalSeq-C Hashtags following manufacturer's directions (BioLegend). Samples were counted, pooled at equal proportions, treated with human TruStain FcX and stained for 30 min with TotalSeq-C Universal Cocktail following manufacturer's directions (BioLegend). Samples for each donor were loaded across two lanes of a 10X Genomics Chromium instrument targeting between 8,000-10,000 cells each. cDNA synthesis and amplification was performed using Next GEM Single Cell 5' Kit v2 (10X Genomics).  In this experiment, the hashtags correspond combinations of tissues and treatment combinations. For D667, HTO1: Unstimulated blood, HTO2: LPS stimulated blood, HTO3: IL4 stimulated blood, HTO4: Unstimulated lung, HTO5: LPS stimulated lung, HTO6: IL4 stimulated lung, HTO7: Unstimulated spleen, HTO8: LPS stimulated spleen, HTO10: IL4 stimulated spleen, HTO12: Unstimulated jejunum, HTO13: LPS stimulated jejunum, HTO14: IL4 stimulated jejunum For D674, HTO2: Unstimulated blood, HTO3: LPS stimulated blood, HTO4: IL4 stimulated blood, HTO5: Unstimulated lung, HTO6: LPS stimulated lung, HTO7: IL4 stimulated lung, HTO8: Unstimulated spleen, HTO9: LPS stimulated spleen, HTO10: IL4 stimulated spleen, HTO12: Unstimulated jejunum, HTO13: LPS stimulated jejunum, HTO14: IL4 stimulated jejunum For D681, HTO2: Unstimulated blood, HTO4: LPS stimulated blood, HTO5: IL4 stimulated blood, HTO6: Unstimulated lung, HTO7: LPS stimulated lung, HTO8: IL4 stimulated lung, HTO9: Unstimulated spleen, HTO10: LPS stimulated spleen, HTO12: IL4 stimulated spleen, HTO13: Unstimulated jejunum, HTO14: LPS stimulated jejunum, HTO15: IL4 stimulated jejunum Sequencing libraries were prepared using Next GEM Single Cell 5' Kit v2 (10X Genomics) and sequenced on a NovaSeq X (Illumina) with 100 cycles for read 1 and 90 cycles for read 2.