Fig. 4.

Activation of de novo RNA, protein, and cytokine synthesis occurs sequentially during ex vivo peripheral human immune activation. a Manually annotated SPADE map of unstimulated, in silico gated, PBMCs clusters, color overlaid with the median baseline incorporation of BRU (left), puromycin (center), and median baseline phosphorylation of 4EBP1-Thr37/46 (right). b A heat map summary, ordered by stimulation time and grouped by cell phenotype, of peripheral immune cell subsets with PMA/ionomycin activation, each parameter normalized to 0 h (ratio of transformed cluster median at given time point relative to t = 0 h). (ArcSinh ratio color scales; p4EBP1, BRU, Puromycin, TNFalpha, IFNgamma: −1 to 1; pS6, CD69: −3 to 3). Number of cell events displayed; 95,000 to 150,000 mononuclear cell events for each time point, experiments were performed multiple times, with representative maps and plots shown