NfκB signaling dynamics quantification in single primary hematopoietic cells. (A) Quantification of nuclear fluorescence dynamics in a representative cell plotted as trace or “heat stripe.” The fading magenta pattern reflects limited TNFα stability after a single addition. (B) Four main NfκB response types upon TNFα challenge. (C) Features extraction from signaling traces for detailed objective analysis (supplemental Table 1). (D) Two-dimensional UMAP representation of cellular heterogeneity across extracted features. One dot represents 1 trace. (E) UMAP as in (D) but with colored dots indicating the localization of cells with a specific response pattern (n = 2721 cells, N = 3 biological replicates). See also supplemental Table 1.