Extended Data Figure 7: JAK–STAT signalling regulates glial lipid metabolism.
From: Gut cytokines modulate olfaction through metabolic reprogramming of glia
a, Workflow of scRNA-seq using plate-based Smart-seq2. FACS, fluorescence-activated cell sorting. Four groups of glia were sequenced: 5 and 50 day all glia (GFP+, driven by rep::Gal4); 5 and 50 day EG (GFP+, driven by GMR56F03::Gal4). b, Visualization of glial cells using t-SNE plots. Cells were coloured according to cell types, ages and Louvain clusters with default resolution. Non-EG were curated from all repo+ glia with EG (GMR56F03::Gal4+) removed (Methods). EG and non-EG were readily separated into different clusters (left and middle). In total, 10 clusters were formed from these glia (right), suggesting the heterogeneity of glial population. c, Violin plot showing expression levels of dome in non-EG and EG. For both EG and non-EG, cells were combined from young and old flies. In non-EG, dome expression was barely detected except in one cell. In EG, a subset of cells showed high expression of dome. d, Violin plots showing expression levels of Socs36E in young and old non-EG (left) and EG (right) respectively. e, Visualization of all annotated glial cells from a previously published whole fly brain scRNA-seq dataset22 using a t-SNE plot. scRNA-seq was performed using droplet-based 10x Genomics platform. Glia are in red (repo+), and neurons are in grey. Two subsets of EG (in orange box) and six subsets of non-EG (in blue box) are annotated. f, Violin plots showing expression levels of Socs36E in non-EG and EG at eight different ages. Cells from 3-, 6- and 9-day-old flies were combined as young samples, and compared with cells from 50-day-old flies (old). g, Gating strategy for sorting STAT::GFP+ glia and STAT::GFP− glia from the central brain of young mock or young infected (4-h Ecc15 infection) flies overexpressing tdTomato in all glia (repo::Gal4) while expressing 10xSTAT::GFP reporter. h, Visualization of gene expression variation between STAT::GFP+ glia and STAT::GFP− glia by PCA plot. Each dot represents a sample replicate independently collected from a cohort of 100 flies. Samples with the same genotype were grouped together, and samples with different treatments were coloured separately. i, Volcano plot displaying differentially expressed genes between STAT::GFP+ glia and STAT::GFP- glia (highlighted in red) under homeostatic conditions, using a cut-off of twofold change, P < 0.001, FDR < 0.01. j, Gene Ontology analysis of significantly upregulated genes in STAT::GFP+ glia during homeostasis. k, Lipid storage-associated genes were significantly upregulated in STAT::GFP+ glia during homeostasis. Reads per kilobase per normalized million mapped reads (nRPKM) values of each gene in STAT::GFP+ glia and STAT::GFP− glia are shown correspondingly. l, Genes involved in monocarboxylate transport were significantly upregulated in STAT::GFP+ glia during homeostasis. nRPKM values of each gene in STAT::GFP+ glia and STAT::GFP− glia are shown correspondingly. m, Schematic demonstrating mitochondrial fatty acid β-oxidation. n, Genes involved in fatty acid β-oxidation that were significantly upregulated in STAT::GFP+ glia during homeostasis. nRPKM values of each gene in STAT::GFP+ glia and STAT::GFP− glia are shown correspondingly. Data are mean and s.e.m. The sample size is as follows: n = 4 replicates per condition (each replicate was independently pooled from 100 flies on different days) in g–n. P values in k, l and n were calculated by Partek Flow; P values in c, d and f from one-tailed Student’s t-test.
