Fig. 5: Comparative transcriptomic effects of FOXO3 and FOXO1 expression.

A Principal component analysis of RNA-sequencing datasets from FACS-sorted replicate 697 cultures transduced with SIN-SIEW, SIN-SIEW-FOXO3 or SIN-SIEW-FOXO1. PC1 – principal component 1; PC2 – principal component 2. B Venn diagram illustrating unique and overlapping gene sets differentially expressed (adjusted p-value < 0.01, log-2 fold-change >1) by forced expression of FOXO3 and FOXO1 in 697 cells. C Heat map showing expression relative to control levels of the genes dysregulated by FOXO1 and FOXO3. The scale has been truncated at log₂-fold-change 2.5 to clearly illustrate the more potent overall effects of FOXO1 expression on transcription levels. Figures A-C were generated in R/RStudio. D Heatmaps, generated in Morpheus, showing relative expression, in replicate 697 cultures transduced with SIN-SIEW or FOXO3 / FOXO1 expression constructs, of genes in MSigDB datasets. The examples shown are illustrative of multiple gene sets showing differential expression of; immunoglobulin heavy (IGHV) and light (IGL) chain genes, small ribosomal subunits (Large ribosomal subunits were also downregulated by FOXO1), HLA class II and metallothionein genes. E Histograms showing mean adjusted RNA read counts (TPM) for specific genes involved in early B-cell development for replicate 697 cultures transduced with SIN-SIEW or FOXO expression constructs. Error bars indicate SEM values, significance is indicated by **p < 0.01, *p < 0.05, (p > 0.05).