Figure 4,F.

The phospho-flow protocol produces reliable signaling results and allows for trainee customization. This figure shows the expected signaling outcome from Protocol 1. In part A, one iteration is shown, with heatmaps indicating the arcsinh transformed ration of median protein phosphorylation compared to the unstimulated condition. Each row represents a cell population as defined in Figure 3, and each column represents a stimulation condition. Five different phosphorylated protein readouts are shown here. In this experiment, the trainee selected peroxide as their third condition. In part B, the results of two trainees performing the protocol in parallel are shown. In part C, a variety of ‘wildcard’ stimuli and their most robust readouts are shown.