Fig 1: Mechanisms for local medulla injection of ADPN promoting bone healing. (A) Serum Osteoprotegerin (OPG) levels at weeks 2, 4, and 6. (B) WB analysis of alkaline phosphatase (ALP), bone morphogenic protein 2 (BMP-2), osteocalcin (OCN), and adiponectin receptor 1 (AdipoR1) expressions with corresponding quantification, (C) Immunofluorescent staining of ALP, OCN, BMP-2, and AdipoR1 in G2 (ADPN 1 mg/kg) and G3 (ADPN 2 mg/kg), and the corresponding quantification, n = 5. The white arrow indicates the periosteum, and the blue arrow indicates the lacuna, scale bar = 100 µm.
Fig 2: Histology evaluation of bone regeneration. The hematoxylin and eosin (H&E) and Masson trichrome staining (MS) images of rat tibia transverse sections at weeks 2, 4, and 6 (n = 5), scale bar = 500 µm and scale bar = 200 µm in the magnified images. The ADPN treatment groups showed thicker cortical bone and bone calluses. The callus thickness and the blue-stained area, indicative of new bone formation, were quantified and plotted.
Fig 3: Activation of APN/CPT signaling alleviates pulmonary fibrosis in BLM-induced IPF rats. An IPF rat model was established by injection of BLM, and the BLM-induced rats were injected with C75 and/or 3-MA for treatment. (a) The histological analysis was conducted by H&E staining. (b) The protein expression level of TGF-ß, a-SMA, and Collagen I in pulmonary tissues was measured using western blot. (c–e) The levels of the lipid peroxides including 4-HNE, MDA, and ox-LDL in pulmonary tissues were detected using their commercial kits, respectively. The level of APN (f) in blood and (g) in pulmonary tissue was measured by ELISA. (h) The protein expression of AdipoR1 and CPT1A in pulmonary tissues was detected by western blot. (i) The protein expression of Beclin-1, Atg5, and LC3B II/I in pulmonary tissues was detected by western blot. ***p < 0.001 vs. blank; #p < 0.05, ##p < 0.01, and ###p < 0.001 vs. BLM; +p < 0.05, ++p < 0.01, and +++p < 0.001 vs. BLM+C75.
Fig 4: Hypoxia induces macrophage M1 phenotype and oxidative stress but lowers autophagy and APN/CPT1A signaling. THP-1 cells were differentiated into M0 macrophages by induction of PMA, followed by hypoxia induction. (a) The level of CD86 and iNOS was detected by immunofluorescence. (b) The LC3B-positive cells were determined using immunofluorescence and observed under a fluorescence microscope. (c) The protein expression of beclin-1, Atg5, and LC3B II/I was measured by western blot. (d) The level of HIF-1a and mTOR was detected by ELISA. (e) The level of lipid peroxides, such as 4-HNE, MDA, ox-LDL, and ROS, was measured using their commercial kits, respectively. (f) The protein expression of AdipoR1 and CPT1A was measured using western blot. ***p < 0.001 vs. M0.
Fig 5: The serum concentrations of adiponectin (ADPN) based on medulla injections, n = 5.
Supplier Page from Abcam for Rat Adiponectin ELISA Kit