Flow cytometry of nuclei populations from the mouse hippocampus after intranuclear stains with inCITE antibodies, followed by Alexa Fluor 647-conjugated secondary stain: NeuN in PBS (a), PU.1 in PBS (b), p65 in kainic acid (KA) (d), and c-Fos in PBS (e) and kainic acid (KA) (f) treated mice. Axes show fluorescence signal (x axis) and side scatter (y axis) of singlet nuclei (dots); histograms show marginal distributions. Oval gates show NeuNhigh (a, 58.3%), PU.1high (b, <3%), p65high (d, 55.2%), c-Foshigh (0.21% in PBS (e), and 48.7% after KA treatment (f)). c. Right: Distribution of PU.1 in microglia (CD11b+ CX3CR1+, red), CD4+ cells (blue) and isotype (gray) cells measured by flow cytometry (left and middle panels) after simultaneous surface protein and intracellular protein stains (Methods).