Fig. 5: Transcriptional changes induced by fasting.

a, Fos is increased in nucSeq AgRP neurons after fasting. Left, UMAP plot depicting C66-46: Agrp.GABA-4 in HypoMap. The inset shows Agrp expression in nucSeq cells from C66-46: Agrp.GABA-4. Right, UMAP plots of the same cells showing Fos expression in fasted and ad-libitum-fed conditions. The changes in nucSeq AgRP neurons after fasting were strong enough to cause a shift in the cluster. b, IEGs with high log₂(fold change) (log₂FC) in AgRP neurons. The violin plots show the per-cell expression between conditions. c, Neuron clusters activated by fasting. The bar plot depicts the percentage of significantly up-regulated IEGs in the fasted state over the total number of expressed IEGs (left number, based on presence in at least 10% cells of clusters in either condition). AgRP neurons are strongly activated, as indicated by the high number of changing IEGs. The bars are colored by mean log₂FC, and the number of cells in each cluster is shown on the right. d, Transcriptional changes in AgRP neurons induced by fasting. In the volcano plot (log₂FC versus adjusted P values from a (two-sided) Wald test), differentially expressed genes (DEGs) are highlighted. The dot plot shows Gene Ontology (GO) terms enriched in up-regulated genes. The P values are based on a hypergeometric test from an over-representation analysis and were corrected using false discovery rate (FDR). e, Comparison of transcriptional changes in AgRP neurons between nucSeq and Campbell et al.⁵ data. The scatter plot of log₂FCs is colored by DEGs in either dataset. f, Per-cell expression levels of selected DEGs between conditions. For each gene, the expression is shown across multiple activated cell types as well as POMC neurons and a reference containing all remaining cells. P values of DEGs were obtained by Wilcoxon rank-sum tests and were adjusted for multiple comparisons using Bonferroni correction. See also Supplementary Tables 10 and 11.

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