Fig. 1: SETD5 associates with NCoR and HDAC to inhibit Cebpa and Pparg induction during adipogenesis. | Nature Communications

Fig. 1: SETD5 associates with NCoR and HDAC to inhibit Cebpa and Pparg induction during adipogenesis.

From: Spatiotemporal dynamics of SETD5-containing NCoR–HDAC3 complex determines enhancer activation for adipogenesis

Fig. 1

a Schematic illustration of enhancer states. Poised enhancer in embryonic stem cells (ESCs) is characterized by the presence of H3K4me1 and H3K27me3. Primed enhancer is characterized by the presence of the only H3K4me1, while active enhancer is characterized by the presence of both H3K4me1 and H3K27ac6. b Schematic representation of mouse SETD5 protein and its mutants. c Retroviral expression of SETD5 in 3T3-L1 preadipocytes. Nuclear proteins from indicated 3T3-L1 preadipocytes were subjected to immunoblot (IB) analysis. Equal loading of the proteins was confirmed by blotting with anti-histone H3 (H3) antibody. d ORO staining of 3T3-L1 preadipocytes transduced with empty virus or SETD5. Preadipocytes were induced with MDI mixture or MDI mixture plus troglitazone (Tro). e Transcriptional changes of adipogenic genes in SETD5-transduced 3T3-L1 preadipocytes during adipogenesis by using a microarray. f, g siRNA-mediated knockdown of SETD5 in 3T3-L1 preadipocytes. Setd5 (f), Cebpa, and Pparg (g) mRNA expression was quantified by qPCR. h, i ORO staining of SETD5 knocked-down 3T3-L1 preadipocytes (h) or 3T3-L1 preadipocytes transduced with empty virus, SETD5, or mutants (i). Preadipocytes were induced with Dex (h), or MDI mixture (i). j Proteomics analysis of SETD5 interacting proteins. Nuclear proteins from WT-, ΔSET-, and Δ437-918-SETD5-transduced preadipocytes were subjected to immunoprecipitation with anti-V5 antibody followed by trypsin digestion and mass spectrometry. Color code: proteins lost (<2−3-fold) by deletion of a.a. 437–918 (blue); proteins lost by deletion of SET domain (orange); proteins not affected by deletion of a.a. 437–918 or SET domain (white). Proteins of PAF1 complex are highlighted in black. Proteins of APC/C complexes and RNF213 are highlighted in pink. k siRNA-mediated knockdown of NCoR2 in SETD5-transduced 3T3-L1 preadipocytes. Ncor2 mRNA expression was quantified by qPCR. l ORO staining of NCoR2 knocked-down, SETD5-transduced 3T3-L1 preadipocytes. Scale bar = 50 μm. c, f, g, k, l Representative of three independent experiments. f, g, k Data are mean ± SD of three technical replicates. f, g One-way ANOVA with Tukey’s multiple comparisons test. k Unpaired two-tailed Student’s t-test. **p < 0.01. Source data are provided as a Source data file.

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