ebf2 flox flox conditional null mice  (Cyagen Biosciences)

Journal: The Journal of Clinical Investigation

Article Title: EBF2 variant identified in a patient with atypical partial lipodystrophy causes adipose fibrosis and dysfunction

doi: 10.1172/JCI192737

Figure Lengend Snippet: ( A ) Photographs of a patient with atypical PLD showing altered fat distribution, a dorsal neck fat pad with acanthosis (arrow), and prominent veins in the legs (arrows). ( B ) Whole-body dual-energy X-ray absorptiometry (DEXA) “fat shadow.” ( C ) H&E- and Masson’s trichrome–stained images of liver, kidney, and subcutaneous WAT (abdomen, neck, thigh); collagen is shown in blue. ( D ) Sanger sequencing showing G>T substitution. ( E ) 3T3-L1 cells transduced with control, full-length EBF2, or truncated EBF2 (1-164). GFP (green); BODIPY (red); DAPI (blue). ( F ) Luciferase reporter assay in HEK cells ( n = 3). Scale bars: 100 μm ( C and E ). **** P < 0.0001, by 1-way ANOVA with Tukey’s test ( F ). Data indicate the mean ± SEM. Each dot represents a biological replicate (see also ).

Article Snippet: Ebf2 E165/X -KI mice were generated on a C57BL/6J background (strain no. 000664, The Jackson Laboratory) using CRISPR/Cas9 technology at the University of Michigan Transgenic Core.

Techniques: Staining, Sequencing, Transduction, Control, Luciferase, Reporter Assay

Journal: The Journal of Clinical Investigation

Article Title: EBF2 variant identified in a patient with atypical partial lipodystrophy causes adipose fibrosis and dysfunction

doi: 10.1172/JCI192737

Figure Lengend Snippet: ( A ) Allele-specific RT-qPCR of IWAT ( n = 3–5). ( B ) Differentiated IWAT SVF from Ebf2 +/+ and Ebf2 E165X/+ . BODIPY (green), DAPI (blue). (C ) Quantification of lipid accumulation ( n = 6). ( D and E ) Expression of adipogenic genes and Ebf family members in the SVF before and after differentiation ( n = 5). ( F ) Effects of truncated and full-length EBF2 and shRNAs in the SVF. ( G and H ) Quantification of lipid accumulation and gene expression ( n = 3). Scale bars: 100 μm ( B and F ). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001, by 1 or 2-way ANOVA with Tukey’s test ( C – E , G , and H ). Data indicate the mean ± SEM. See also .

Article Snippet: Ebf2 E165/X -KI mice were generated on a C57BL/6J background (strain no. 000664, The Jackson Laboratory) using CRISPR/Cas9 technology at the University of Michigan Transgenic Core.

Techniques: Quantitative RT-PCR, Expressing, Gene Expression

Journal: The Journal of Clinical Investigation

Article Title: EBF2 variant identified in a patient with atypical partial lipodystrophy causes adipose fibrosis and dysfunction

doi: 10.1172/JCI192737

Figure Lengend Snippet: ( A ) Human subcutaneous fat-derived preadipocytes were transduced with control, full-length, or truncated EBF2 . BODIPY (red), GFP (green), DAPI (blue). ( B ) BODIPY quantification ( n = 2). ( C ) Venn diagram of RNA-seq of differentially expressed genes. WT, EBF2; Mut, EBF2 (1-164). Mut, mutation. ( D and E ) Heatmaps of ECM and cytokine receptor genes. ( F and G ) CyTOF images of the patient’s neck WAT showing DNA, collagen I, CD34, CD15, CD11c, and CD68. ( H and I ) Immunofluorescence images of patient’s and control WAT depots. Scale bars: 100 μm ( A and F – I ). ** P < 0.01, by 1-way ANOVA with Tukey’s test ( B ). Data indicate the mean ± SEM. Each dot represents a single image.

Article Snippet: Ebf2 E165/X -KI mice were generated on a C57BL/6J background (strain no. 000664, The Jackson Laboratory) using CRISPR/Cas9 technology at the University of Michigan Transgenic Core.

Techniques: Derivative Assay, Transduction, Control, RNA Sequencing, Mutagenesis, Immunofluorescence

Journal: Cell reports

Article Title: Defective brown adipose tissue thermogenesis and impaired glucose metabolism in mice lacking Letmd1

doi: 10.1016/j.celrep.2021.110104

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Primary antibodies used in this study were anti-Letmd1 (LSBio; LS-C335200), anti-Brg1 (Bethyl; A300-813A-T or Cell Signaling; 49360S), anti-Ucp1 (R&D systems; MAB6158), anti-Prdm16 (R&D systems; AF6295), anti-Ebf2 (R&D systems; AF7006), anti-OXPHOS (MitoSciences; MS604), anti-lamin (Santa Cruz Biotechnology; sc-376248), anti-VDAC (Cell Signaling; 4661T), anti-FABP4 (Santa Cruz; sc-271529), anti-Flag (GenScript; A00187), anti-V5 (ThermoFisher; R96025), anti-β-actin (Santa Cruz; sc-47778), anti-Gapdh (Santa Cruz; sc-25778), and anti-vinculin (Sigma, V9131).

Techniques: Recombinant, Cell Culture, Reverse Transcription, SYBR Green Assay, Chromatin Immunoprecipitation, Plasmid Preparation, Software