A, Uniform manifold approximation and projection (UMAP) dimensional
reduction of wild-type (WT)-corn oil (CO) (8037 spots, 3 sections), WT-tamoxifen
(TAM; 8882 spots, 3 sections),
PSAPP/Inpp5dfl/fl/Cx3xr1CreER+
(PSAPP)-CO (7748 spots, 3 sections), and PSAPP-TAM (9366 spots, 3 sections).
Each dot represents a Visium transcriptomics spot. The 27 color-coded clusters
correspond to brain areas/regions (see below). B, Chord diagram of genes
enriched > 1 log2 fold (l2f) in each cluster and adjusted
P-value < 0.05. Each string represents a gene shared
between two and six other clusters. C, Example clusters highlighted in a Visium
brain section. Boxes show top three enriched genes per cluster. Underlined gene
is highlighted in the SpatialFeaturePlot. D, UMAP with Cluster 26 highlighted
showing that it is only present in PSAPP animals. E, Volcano plot of genes
enriched in Cluster 26 as calculated with Seurat’s
FindAllMarkers function. Red genes are l2f > 1 and padj < 0.05,
blue l2f < 1 and padj < 0.05, green genes are l2f > 1 and
padj > 0.05, gray genes are l2f < 1 and padj > 0.05. F,
Module score of Cluster 26 and 6E10 (amyloid) staining of the adjacent brain
section shows colocalization. f’ shows higher magnification of box in
(F). G, Gene Ontology term analysis of genes enriched l2f > 1 in Cluster
26 using metascape.73 H, UpSet
plot highlighting the overlapping and unique genes between previously defined
reactive microglia and astrocyte subtypes: Disease-associated microglia, DAM1
(early) and DAM2 (late),25
interferon-responsive reactive astrocytes (IRRAs),28 plaque-induced genes (PIGs),27 as well as disease associated
astrocytes (DAA).26 Insets are
module scores of the genes in each reactive subtype highlighted on a PSAPP-TAM
spatial transcriptomics brain section. For IRRAs, DAAs and Cluster 26, genes
were filtered for padj < 0.05 and l2f > 0.5. For DAM1, DAM2, and
PIGs, genes were selected based on gene sets provided in the original
publication. I, Genes differentially expressed in Cluster 26 between APP-CO and
APP-TAM as calculated with edgeR on sum of counts using
muscat (red genes are l2f > 1 and padj <
0.05, blue l2f < 1 and padj < 0.05, green genes are l2f > 1
and padj > 0.05, gray genes are l2f < 1 and padj >
0.05).