(a) Smoothed representation of statistical tSNE map (show in Supplementary Fig. 9b) after thresholding to 0.05 FDR-adjusted p-values (blue) shows selection of four differentially abundant subsets by automated gating (black contour lines). Skilling-Mack testing was performed on n = 35 biologically independent huMG samples (SVZ (n = 8); THA (n = 8); CER (n = 5); GTS (n = 8) and GFM (n =6)) from 8 biologically independent donors. (b) Snail plot shows mean marker expression levels of each huMG subset (subset 1 = red; 2 = orange; 3 = green and 4 = purple). The snail shell represents transversal (perpendicular) axis mapping marker expression levels on exponential scale. Each line denotes each sample (total of 36 samples). (c) Gates of four detected subsets overlaid on cell density tSNE plots of concatenated FCS files for each brain region (biologically independent samples of SVZ = 8; THA = 8; CER = 5; GTS = 8 and GFM = 7). (d) Top panel shows median bin-expression levels in boxplot representation of markers that discriminate between the four subsets (subset 1 = red box (n = 6 bins); 2 = orange box plot (n = 95 bins); 3 = green box (n = 4 bins); 4 = purple box (n = 12 bins); remaining cells (cells – subsets) = blue box (n = 395 bins) according to robust separation score. Box center line and limits represent median, 16th and 84th percentiles; whiskers define the data minimum and maximum.