Transcriptional differences between fCD8s and GCTfh’s. (A) Analysis showing a significant increase in BCL6 mean fluorescence intensity (MFI) in GCTfh’s compared with fCD8s and non-fCD8s (n = 5). (B) PCA of RNA-Seq data from the 4 cell subsets. The top 500 differentially expressed genes were used to construct the PCA plot. Clear separations are observed between the subsets, with the fCD8 and non-fCD8 subsets showing the closest proximity. (C) Statistical analysis showing the significantly greater magnitude of BCL6 expression in GCTfh’s compared with fCD8s and no difference between fCD8s and non-fCD8s (n = 5). (D) Heat map showing the relative expression of the indicated genes. Genes were ranked from highest expression (red) to lowest expression (blue). Each column represents the expression level for a particular patient as labeled on the x-axis.