Supplementary Figure 5: Communication network assembly and validation of novel connections.

(a) Relative coverage of immune annotated transcription factors (T), adaptor molecules (A), receptors (R), and ligands (L).

(b) Histogram of intercellular interaction scores. Intercellular interaction scores of our study were reduced to cell types and cytokines identified in the textbook (light grey). Intercellular interaction scores covered by the textbook are displayed in (dark grey).

(c) Intercellular interaction score cutoff (Online Methods).

(d) Percentage of ingoing connections for cytokines in our study compared to ‘Textbook’ and ‘ImmuneXpresso’.

(e) Understudied cell types and cytokines from ingoing connections. Significantly enriched cell types and cytokines in our study compared to ‘Textbook’ and ‘ImmuneXpresso’ were identified by Fisher’s exact test with BH FDR of 5%.

(f) Resistin (RETN) secretion of B cells. B cells were stimulated with a cocktail (Goat F(ab’)2 anti-human Lambda/Kappa, F(ab’)2 Goat anti-mouse IgG Fc, CpG, anti-CD40) or LPS and anti-CD40 for 4 days and the resistin concentration in the supernatant was determined by ELISA (n=4, cell culture replicates from 2 independent donors).

(g) T4 memory cells respond to IL-34 by total proteome changes. T4 memory cells were activated with aCD3/aCD28 for 2 days and then cultured for another 2 days with IL-2 in the presence of different amounts of IL-34 (0, 1, 10, or 100 ng/ml). The volcano plot (top) shows the protein differences between T4 memory cells treated with 0 ng/ml and 100 ng/ml IL-34 (significantly changed proteins are shown in blue, S₀=1, FDR < 5%, n=4, cell culture replicates from 2 independent donors). Examples for significantly shifted Keyword annotations (determined by 1D annotation enrichment, BH FDR < 1%) are highlighted in violet (calcium transport) or green (immunglobulinVregion). Density distribution (bottom) show the shift of the Keyword annotations in dependency of IL-34 concentration.