Figure 4.

High-Content Profiling of Single-Cell Responses to Dynamic Immune Inputs

(A) Top: fluorescence and brightfield image series of a single cell over the course of LPS stimulation experiment. The scale bar represents 20 μm. Bottom: corresponding data from this cell for NF-κB (RelA) activity, TNF secretion, morphology, and migration responses are shown. LPS stimulation depicted by the tan bar was 500 ng ml⁻¹ for 2 hr.

(B) RelA translocation and TNF secretion for distinct LPS input dynamics. The single 2-hr pulse of LPS (left), continuous LPS exposure (center), and 8-min pulses of LPS every 2 hr (right) are shown. The bars above heatmaps depict stimulation profiles, and scale bars depict normalized nuclear RelA and molecules of TNF. Continuous stimulation leads to significantly increased cell-to-cell variability and oscillatory dynamics in TNF secretion compared to transient or pulsed stimulation. RelA is normalized to initial nuclear fluorescence intensity of each cell prior to stimulation.