Fig. 5: Single-cell spatial proteome analysis of the cerebellar cortex. | Communications Biology

Fig. 5: Single-cell spatial proteome analysis of the cerebellar cortex.

From: Maternal immune activation followed by peripubertal stress combinedly produce reactive microglia and confine cerebellar cognition

Fig. 5

a Classification of cell populations from IMC data. Single-cell segmentation superimposed with the IMC images of male 2HIT susceptible cerebellar cortex. b Heat map of the marker signal intensity by clusters. c tSNE plot and annotation of the clusters: immune cells, granule cell layer (GCL) neurons, molecular layer interneurons (MLIs), vasculature cells, astrocytes, oligodendrocytes, and unknown populations. In 2HIT, we termed cluster 6 progenitors of stress-associated microglia/macrophage (proSAM), cluster 12 SAM, and cluster 14 border-associated macrophage (BAM). BAM was annotated from the expression of Lyve1, MHC class II, and CX3CR1, and localization in the tissue. See also Supplementary Fig. 12. d Intensity plot of representative molecules in the tSNE maps. e tSNE plot of the mixture dataset of Control and 2HIT. The successful batch correction has effectively integrated the cells into the same clusters from both models, suggesting similar cell populations in the overall dataset. f Volcano plot showing the changes in the protein expression between immune cells and SAM cluster of the mixture dataset. Red-marked proteins have significantly higher expression in SAM compared to all the other immune cells. g Representative SAM images by immunohistochemistry, showing Iba1-, MHC class II-, TREM2-triple-positive-microglia in male 2HIT cerebellum. The density of MHC class II(+) immune cell (h) and Iba1-, MHC class II-, TREM2-triple-positive-cell (i) of cerebellar cortex (CbCrx), cerebellar nuclei (CbN), hippocampus (HPC), and medial prefrontal cortex (mPFC) in male Control, 2HIT sus, 2HIT res, and 2HIT+rMG. *p < 0.05, one-way ANOVA with multiple comparisons. j Representative SAM images by immunohistochemistry in female 2HIT cerebellum. The density of MHC class II(+) immune cell (k) and Iba1-, MHC class II-, TREM2-triple-positive-cell (l) in female Control, 2HIT sus, 2HIT res, and 2HIT+rMG. *p < 0.05, one-way ANOVA with multiple comparisons using the Tukey-Kramer method.

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