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Journal: iScience
Article Title: Intercellular cGAMP transmission induces innate immune activation and tissue inflammation in Trex1 deficiency
doi: 10.1016/j.isci.2021.102833
Figure Lengend Snippet: Delivery of cGAMP from radioresistant cells to immune cells induces tonic IFN and NF-κB signaling in Trex1 deficiency (A) Global gene expression analysis by 3′-mRNA seq. Gene and sample-wise hierarchical clustering based on differentially expressed genes (non-adjusted p value <0.05, n = 660) within the dataset of spleen tissue from Cgas −/− BM and dKO BM mice. Gene expression values are Z score standardized. For visualization purposes Z scores were limited to numbers between −2 and 2. (B) PCA plot of spleen tissue data from Cgas −/− BM and dKO BM mice. PCA is based on differentially expressed genes (non-adjusted p value <0.05, n = 684). (C) Volcano plot indicating transcriptomic changes between Cgas −/− BM and dKO BM mice. Genes with a non-adjusted p value <0.05 and a signed fold change >2 are colored in red (upregulated in Cgas −/− KO BM mice) and blue (upregulated in dKO BM mice). Differentially expressed genes with an adjusted p value < 0.05 are labeled. (D) Normalized Enrichment Scores (NES) of Hallmark gene sets significantly upregulated (FDR q value <0.01) in spleen tissue from Cgas −/− BM mice compared with that from dKO BM mice in a pre-ranked Gene Set Enrichment Analysis (GSEA) based on a metric score calculated by log 2 (fold change) x (-log 10 (p value)). n = 5 mice per group. (E) Percentage of splenic classical monocytes and red pulp macrophages with high expression of IκBα (n = 7 for Cgas −/− BM mice, n = 6 for dKO BM mice). (F) Fold change of p-RelA mean fluorescence intensity (MFI) of classical monocytes and red pulp macrophages in instantly fixed spleens from Cgas −/− BM (n = 7) and dKO BM (n = 6) mice. Individual fold change values were calculated as ΔMFI i = (MFI i , untreated – MFI i , λPP ) divided by the average of ΔMFI i values obtained from dKO BM mice. (G) Histograms display representative results for IκBα staining of gated classical monocytes and red pulp macrophages. Data in bar graphs are represented as mean + SEM. Data in (E-G) are pooled from two independent experiments. Statistically significant differences were determined by two-tailed unpaired Mann-Whitney U test ( ∗ p < 0.05, ∗∗ p < 0.01). See also Figure S2 .
Article Snippet: For gene set enrichment analysis (GSEA) a pre-ranked list was generated based on a metric score calculated by: l o g 2 ( f o l d c h a n g e ) x ( − l o g 10 ( p v a l u e ) ) and applied to the GSEA java desktop application (version 4.0.3) provided by the Broad Institute ( ).
Techniques: Gene Expression, Labeling, Expressing, Fluorescence, Staining, Two Tailed Test, MANN-WHITNEY