Heat map analysis of flow cytometric data for identification of pathway inhibitors. Screenshots of WebFlow heat maps generated from the sample U937 stimulation and inhibition experiment. U937 cells were treated with a dose–response of Jak inhibitor I (a pan-Jak kinase inhibitor) in rows C and F prior to stimulation with IFN-γ to induce Stat1 phosphorylation. Phosphorylation of Stat1 was measured by staining with an antibody against pStat1 (Y701). Demonstrated is the ability to change which statistic is represented by the heat map colors, and which is represented by the numbers on the heat map. (A) Heat map colored by median fluorescence intensity (MFI) in phospho-Stat1 channel, and numbers display pStat1 MFI values. (B) Heat map colored by median staining in pStat1, while numbers display Jak inhibitor I concentration. (C) Heat map colored by fold change in pStat1 relative to column 1, and numbers display those values. (D) Heat map colored by percentage inhibition of induced pStat1 levels relative to negative control in column 1 and positive control in column 12, and numbers display those values. In each case, the range of the heat map coloring was adjusted to reflect the range of the data for the calculated statistic.