Journal: Molecular Metabolism

Article Title: Critical changes in hypothalamic gene networks in response to pancreatic cancer as found by single-cell RNA sequencing

doi: 10.1016/j.molmet.2022.101441

Figure Lengend Snippet: MBH endothelial cell response to pancreatic cancer . Differentially expressed (DE) genes between sham and TB mice in endothelial cells as shown with a volcano plot ( A ). Featureplots showing the induction of genes in TB mice linked to the immune system ( Lcn2 , Lrg1 ), membrane organization ( Scgb3a1 ), as well as secreted protein Wnt5a ( B ). Vln plots showing examples of highly upregulated DE genes, highly downregulated DE genes as well as conserved genes between sham and TB mice in the endothelial cluster ( C ). Top 24 upregulated genes in endothelial cells in TB mice sorted by fold inductions ( D ). qPCR analysis of top-induced genes using whole hypothalamic RNA extracts from sham and TB mice ( E ). IHC for ICAM-1 and PECAM1 in the MBH for sham and TB mice ( F ). PECAM1 labels all endothelial cells, while ICAM-1 is only expressed in a subset of the endothelial cells. Co-expression network using the top-induced genes in Lcn2 + endothelial cells revealing a network linked to inflammation. Z-scores indicate REACTOME pathway enrichment for interleukin signaling ( G ). Reactome pathway analysis of top 60 induced genes in endothelial cells of TB mice ( H ). qPCR values represent the mean ± SEM of at least three independent experiments and statistical significance between groups was determined with a Student's t-test (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001). Scale bar = 100 μm.

Article Snippet: IHC for the 30 μm sections was performed by incubating them with commercial antibodies for CD45 (BD, rat, 30-F11) and IBA1 (Wako, NCNP24) or PECAM1 (BD, 550274) and ICAM-1 (R&D, AF796) in blocking buffer (PBS + 5% donkey serum + 0.3% Triton X-100) overnight at 4 °C.

Techniques: Expressing