Fig. 6: Loss of miR-33 in AgRP neurons promotes changes associated with increased activity and mitochondrial biogenesis/function.

a Schematic depiction of approach used for performing scRNA-seq in cells isolated from the hypothalamus of wildtype (WT) and miR-33^AgRPiKO mice under fed condition after high fat diet (HFD) feeding. Generated using BioRender.com. b tSNE plot of cell clusters identified from SC-seq analysis. c UMAP projection of single cell profiles from WT (red) and miR-33^AgRPiKO (blue) mice of AgRP⁺ positive neurons identified from SC-seq analysis and violin plots of AgRP expression in AgRP⁺ neurons from WT and miR-33^AgRPiKO mice. d Canonical pathways represented by Z-score among differentially expressed genes in scRNA-seq analysis of AgRP neurons from WT and miR-33^AgRPiKO mice. Red bars indicate pathways in which genes are consistently upregulated, and blue downregulated based on the predicted Z-score. All represented pathways were significantly changed with a −Log P-value > 1.3. e Dot plots representing the percentage of AgRP neurons expressing miR-33 target genes and relative expression within these cells. Red arrows indicate differences in the percentage of cells expressing a gene. f Violin plots and Dot plots representing the percentage of AgRP neurons expressing miR-33 target genes related to AgRP neuronal activity. Red arrows indicate differences in the percentage of cells expressing a gene. g Representative images and quantification of immunofluorescent staining of cFOS (green) in tdTomato + (red) AgRP neurons (n = 5 WT and 3 miR-33^AgRPiKO). Data represents mean +/- SEM. *p < 0.05 as assessed by unpaired 2-sided Student t-test. h Venn diagram of overlap between upregulated genes in AgRP neurons lacking miR-33 and the top 1000 predicted miR-33 target genes from TargetScan7.2. Table including a list genes found to be relevant with AgRP functions. Source data are provided as a Source Data file. a–g (n = 8 WT and 6 miR-33^AgRPiKO).