Fig. 2: Alterations in energy availability impact the chromatin landscape of AgRP neurons.

a Experimental schematic. ARCs from fed, fasted, and leptin-treated NuTRAP^AgRP mice were isolated followed by nuclear isolation. 10,648 ± 1489 nuclei, from 4 mice, were pooled for each condition (fed, fasted, leptin-treated) before being subjected to ATAC-seq. b FACS plot of single nuclei with FITC (GFP) fluorescence on the x-axis and mCherry fluorescence on the y-axis, with filtering gates showing discrete populations for NuTRAP^AgRP GFP+ and GFP- nuclei. c Heatmap showing differentially expressed OCRs for each of the treatment conditions clustered into one of eight patterns (rows). Amplitude of each peak center (±5 kb) is represented in black as indicated. d Volcano plots showing differentially expressed OCRs in fasted vs. fed mice (left, n = 2, 2) or leptin vs. fasted mice (right, n = 2, 2). Red dots correspond to significantly different gained-open and gained-closed regions [fold change >0.5 up (red) or down (red), FDR  <  0.05]; CPM, counts per million. Peaks with a CPM < 1 were not included in volcano plot. FDR-adjusted p-values with a threshold of 0.05 were generated using the Benjamini-Hochberg method to account for multiple comparisons. e Bar graphs revealing the genomic features of fasted-opened, fasted-closed, leptin-opened, leptin-closed ATAC-seq peaks. f Genome browser views (IGV) of ATAC-seq peaks within the vicinity of the Npy gene. Yellow bars correspond to fasted-opened ATAC-peaks. g Genome browser views (IGV) of representative Fasted-opened, Fasted-closed, Leptin-opened, and Leptin-closed ATAC-seq peaks near the indicated genes. The data from ATAC-seq experiments was generated from two independent biological replicates for fed mice, two independent biological replicates for fasted mice, and two independent biological replicates for leptin-treated mice.