P65 dynamics predict GMP colony behavior. (A) Comparison of TNFα and IL1β as stimulating agents. (B) Pedigree and example images of a selected colony. Cells were tracked until generation 4 or higher. Imaging frequency 9 minutes (0-12 hours) and 30 minutes (12-60 hours). Calculation of fate readouts: supplemental Table 2. (C) Correlation of dynamics features with fate readouts. “Oscillation and responsiveness”: features associated with OSC and higher amplitude after stimulation; “trace volatility and noise”: features associated with general fluctuations in p65 signaling activity (n = 323 cells, N = 3 biological replicates). (D) Correlation of selected dynamics features with selected fate readouts. One dot represents 1 colony. Blue line = linear regression model, R = Pearson correlation. OSC correlates with increased future CD115 protein expression and a smaller nuclear area. (E) Selected fate readouts vs stimulating cytokine, cell type, and p65 response dynamics. CD115 and CD24 expressions are observed between 12 and 24 hours after the start of culture.