Fig. 1: LSD1 interacts with GR to control target gene expression in mouse myofibers at physiological glucocorticoid levels.

a Representative immunofluorescent detection of LSD1 (green) and GR (red) in gastrocnemius muscles of 9-week-old wild-type mice. Nuclei were stained with DAPI. A zoomed-in view of the confocal observation is shown on the top right panel. Scale bar, 50 µm. N = 3. b Representative western blot analysis of GR and LSD1 co-immunoprecipitation in gastrocnemius muscle nuclear extracts. Rabbit IgG served as a control for immunoprecipitation. N = 3 mice. Note that the observed discrepancies in the molecular weight on membranes decorated with anti-LSD1 antibody originate from the high sensitivity of LSD1 to salt and/or pH composition of the elution buffer. c, d Tag density map of LSD1 and GR in skeletal muscles, +/− 5 kb from the LSD1 (c) or the GR (d) peak center, and corresponding average tag density profiles. e Localization of GR and LSD1 at the Ddit4 locus. The four GR binding sites located at enhancer (GBSe1, GBSe2 and GBSe3) and promoter (GBSp1) regions are boxed in orange. f Two-step chromatin immunoprecipitation performed with indicated antibodies followed by qPCR analysis (ChIP-reChIP-qPCR) in gastrocnemius muscles of wild-type mice at GBSe1, GBSe2, GBSe3 and GBSp1 of Ddit4. N = 6 biological replicates. Mean ± SEM. Two-way ANOVA with Tukey correction. g Heatmap with hierarchical clustering depicting the mean centered normalized expression of genes differentially expressed in RNA-seq in gastrocnemius muscles of 9-week-old Ctrl, GR^(i)skm-/- and LSD1^skm-/- mice. h, i Pathway analysis of genes down- (h) and up-regulated (i) in gastrocnemius muscles of both GR^(i)skm-/- and LSD1^skm-/- mice at 9 weeks, with p values adjusted using the Benjamini-Hochberg. Source data are provided as a Source Data file.