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. Author manuscript; available in PMC: 2022 Nov 5.
Published in final edited form as: Nat Cell Biol. 2022 May 5;24(5):685–696. doi: 10.1038/s41556-022-00906-y

Fig. 5: Corticosteroid signals regulate distant cebpd expression during heart regeneration.

Fig. 5:

a, Browser tracks of brain ATAC-seq before and after CM ablation, and H3K27Ac and H3K4me3 occupancy of adult brain, kidney, heart muscle and testes. CEN (dashed lines) is open and associated with high levels of H3K27Ac and low levels of H3K4me3 in tissues other than testes. Three biologically independent samples are included for ATAC-seq, and two for ChIP-seq. b, Whole-genome bisulfite sequencing of samples from adult tissues indicating low methylation levels in CEN and cebpd gene regions, with highly methylation in testes. One biological sample for each group. c, Hi-C contact matrix and virtual 4C using the cebpd promoter (top) and CEN (bottom) as bait regions, indicating interactions between CEN and the cebpd gene region in brain. Hi-C data from two biological samples were pooled for each group. d, Footprint analysis reveals enriched binding for NR3C2, AR, and NR3C1 during cardiac regeneration. e, Lollipop plot illustrating predicted NR3C1 and NR3C2 binding sites near cebpd and CEN. f, Schematic of experiments assessing cebpd expression by ISH after agonist treatment. g, Schematic of experiments assessing cebpd expression by qPCR after cardiac injury and antagonist treatment. h, ISH of cebpd on brain (top), head kidney (middle) and heart (bottom) sections after agonist treatment. cebpd is consistently induced in brain and kidney, and only in occasional cases in heart by spironolactone. For brain and head kidney ISH, n = 19 for vehicle, 21 for dexamethasone, 25 for spironolactone. For heart ISH, n = 15 animals for each treatment. Eight of 15 spironolactone-treated animals displayed induction of cebpd expression in heart. Black arrows indicate violet ISH signals. Dex: dexamethasone, Spir: spironolactone. PVZ: periventricular grey zone, Vamgra: the granular layer of the medial division of valvula cerebelli, TL: torus longitudinalis. Black arrows indicate violet ISH signals. Scale bars: 100 μm. i, Quantitative PCR indicating lower cebpd RNA levels in brain after eplerenone, or RU486 plus eplerenone, treatments during heart regeneration. n = 12 animals for DMSO treatment and n = 10 animals for other groups. EPL: eplerenone. Mean ± S.E.M., two-tailed Mann Whitney test.