Journal: bioRxiv

Article Title: Targeting adipocyte ESRRA promotes osteogenesis and vascular formation in adipocyte-rich bone marrow

doi: 10.1101/2023.08.14.552932

Figure Lengend Snippet: (A) Schematic diagram illustrating the experimental procedure for diet-induced obesity (DIO) mice model. 9-week-old Esrra fl/fl and Esrra AKO male mice were fed a high-fat diet (HFD) for 16 weeks. (B) Representative pictures and body weights of DIO mice. (C) Representative images and weight analysis of white adipose tissue (WAT) depots, including gonadal WAT (gWAT), inguinal WAT (iWAT), and mesentery WAT (mWAT). (D) H&E-stained gWAT sections (scale bar: 50 μm). The areas of adipocytes size are presented as graphs. (E-F) Plasma TG (E) and FFA (F) levels. (G) Oral glucose tolerance test (OGTT) was analyzed in Esrra fl/fl and Esrra AKO mice fed a HFD for 12 weeks. (H) Plasma LEPTIN levels. (I) Representative micro-CT images of the distal femoral trabecular bone. (J) Quantitative analysis of bone volume/tissue volume ratio (BV/TV), trabecular thickness (Tb. Th), trabecular number (Tb. N) and trabecular separation (Tb. Sp). (K) H&E staining of femur sections (scale bar: 100 μm). Yellow arrows indicate the bone marrow adipocytes. Osteoblast surface to bone surface ratio (Ob.S/BS) and osteoblast number to bone surface ratio (Ob.N/BS) are shown on the right panel. (L) Calcein double labeling of trabecular bone (scale bar: 20 μm). Mineral apposition rate (MAR) and bone formation rate (BFR/BS) were determined. White arrows indicate the distance between calcein double labeling. (M) TRAP staining of femur sections with quantitative analysis of Oc.S/BS and Oc.N/BS. TRAP-positive purple spots indicate multinucleated osteoclasts (scale bar: 100 μm). (N-O) Plasma P1NP (N) and CTX1 (O) levels. (P) PLIN1 positive marrow adipocytes (PLIN1 + , red) and SPP1 (green) immunofluorescence staining in femur sections (scale bar: 100 μm). The box in the upper showing the metaphysis region near growth plate is represented at higher magnification in the bottom (scale bar: 50 μm). The numbers and areas of adipocytes in the femur marrow per tissue area and the quantification of SPP1 fluorescence intensity were measured. Data are shown as mean ± SD (n = 6 mice). * P < 0.05 was considered statistically significant.

Article Snippet: The paraffin-embedded femurs were subjected to deparaffinization and rehydration procedures before being stained using hematoxylin and eosin (H&E) kit (Beyotime #C0107) or tartrate-resistant acid phosphatase (TRAP) staining kit (Solarbio #G1492).

Techniques: Staining, Micro-CT, Labeling, Immunofluorescence, Fluorescence

Journal: bioRxiv

Article Title: Targeting adipocyte ESRRA promotes osteogenesis and vascular formation in adipocyte-rich bone marrow

doi: 10.1101/2023.08.14.552932

Figure Lengend Snippet: (A) Schematic diagram illustrating the experimental procedure for ovariectomy (OVX)-induced osteoporosis mice model. 10-week-old Esrra fl/fl and Esrra AKO female mice underwent either sham or OVX operation for 8 weeks. (B) Representative images and weights of adipose depots. (C) Representative images and adipocytes size analysis from H&E-stained gWAT sections (scale bar: 50 μm). (D) Plasma LEPTIN levels. (E-F) Micro-CT images of distal femurs in sham and OVX mice (E) with morphometric analysis of BV/TV, Tb.N, Tb.Th, and Tb.Sp (F). (G) Representative TRAP-stained images and quantification of Oc.S/BS and Oc.N/BS in distal femoral metaphysis regions from sham and OVX mice (scale bar: 100 μm). (H) Representative H&E-stained images and quantification of Ob.S/BS and Ob.N/BS (scale bar: 100 μm). (I-J) Plasma P1NP (I) and CTX1 (J) levels. (K) Calcein double labeling with quantitative analysis of MAR and BFR/BS (scale bar: 20 μm). (L) Immunofluorescence co-staining and quantification of PLIN1 + bone marrow adipocytes (red) and SPP1 (green) of femur sections from OVX mice. Scale bar: upper panel, 100 μm; lower panel, 50 μm. Data are shown as mean ± SD (n = 7 mice). * P <0.05 was considered statistically significant.

Article Snippet: The paraffin-embedded femurs were subjected to deparaffinization and rehydration procedures before being stained using hematoxylin and eosin (H&E) kit (Beyotime #C0107) or tartrate-resistant acid phosphatase (TRAP) staining kit (Solarbio #G1492).

Techniques: Staining, Micro-CT, Labeling, Immunofluorescence