ScRNA-seq Analysis of ZsGreen+ Tfh, ZsGreen– Tfh and B cells disassociated from ZsGreen–/+ Tfh and B conjugates

This experiment was designed to study the functional differences between the Th17-derived Tfh and regular Tfh cells. We were also interested in studying how B cells might display functional differences upon receiving help from Th17-derived Tfh cells vs. regular Tfh cells. To achieve these goals, we performed flow sorting and enriched T-B cell doublets with either Th17-derived Tfh or regular Tfh cells to catch the T-B cell interaction “in action”. We then broke the doublets apart with ETDA before scRNA seq analysis to compare the transcriptomic profiles from both T cell side and B cells side. Splenic ZsGreen+ and ZsGreen– Tfh cells conjugated with B cells from K/BxN Th17 fate-mapping mice were both sorted as doublets and then dissociated with EDTA for single-cell RNA sequencing (scRNA-seq) analysis.