Journal: Chinese Medical Journal

Article Title: Impaired autophagy activity-induced abnormal differentiation of bone marrow stem cells is related to adolescent idiopathic scoliosis osteopenia

doi: 10.1097/CM9.0000000000002165

Figure Lengend Snippet: BMSCs from patients with AIS-related osteopenia have a lower osteogenic ability. (A) BMSCs were subjected to osteogenic induction for 21 days. Mineralized nodules were quantified by calculating the ratio of the red mineralization area to the total area after alizarin red staining ( n = 6). Scale bar, 1000 μm. (B, C) The expression levels of BMP2 and COL1 were detected by qPCR after osteogenic induction ( n = 6). (D, E) BMSCs were subjected to osteogenic induction and analyzed over time (3, 7, 10, 14, 17, and 21 days). BMP2 and COL1 were detected by qPCR after 3 weeks of induction. (F–H) BMP2 and COL1 were detected by Western blotting after induction. All data are shown as the mean ± standard deviation. ∗ P < 0.001. AIS: Adolescent idiopathic scoliosis; BMSCs: Bone marrow stem cells; qPCR: Quantitative polymerase chain reaction.

Article Snippet: Primary antibodies against BMP2 (Proteintech, 18933-1-AP, USA), COL1 (Proteintech, 14695-1-AP), C/EBPα (Santa Cruz, sc-365318, China), FABP4 (Santa Cruz, sc-271529 SAMPLE), P62 (Abcam, ab109012, USA), LC3B (Abcam, ab192890), and Beclin 1 (Abcam, ab207612) were used.

Techniques: Staining, Expressing, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Journal: Journal of Translational Medicine

Article Title: Circular RNA EPHA3 suppresses progression and metastasis in prostate cancer through the miR-513a-3p/BMP2 axis

doi: 10.1186/s12967-023-04132-4

Figure Lengend Snippet: BMP2 is the downstream target of miR-513a-3p and promotes proliferation, migration and invasion of PCa cells in vitro. A, B The western blot showed that miR-513a-3p could down-regulate BMP2 protein expression in PC-3 and DU145 cells. C The schematic of BMP2 wild-type and mutant luciferase reporter vectors. D Relative luciferase activity of HEK-293T cells co-transfected with miR-513a-3p mimics or mimics NC, pmirGlo-wild type BMP2 (BMP2-wt) or pmirGlo-mutant type BMP2 (BMP2-mut) plasmids. E, F The proliferation ability of PC-3 and DU145 cells transfected with BMP2 siRNAs or negative control was evaluated by CCK-8 assay. G, I The migration capability of PC-3 and DU145 cells transfected with BMP2 siRNAs or negative control was measured by wound healing assays. Scale bars = 100 μm. H, J–K The migration and invasion capabilities of PC-3 and DU145 cells transfected with BMP2 siRNAs or negative control were assessed by transwell migration and invasion assays. Scale bars = 100 μm. *, ** and *** represented P < 0.05, 0.01 and 0.001, respectively

Article Snippet: The primary antibodies against BMP2 (A0231) and β-actin (AC026) were purchased from ABclonal Biotechnology Co., Ltd. (China), and the secondary goat anti-rabbit antibody (BA1055) was procured from Boster Biotechnology Co., Ltd. (China).

Techniques: Migration, In Vitro, Western Blot, Expressing, Mutagenesis, Luciferase, Activity Assay, Transfection, Negative Control, CCK-8 Assay

Journal: Journal of Translational Medicine

Article Title: Circular RNA EPHA3 suppresses progression and metastasis in prostate cancer through the miR-513a-3p/BMP2 axis

doi: 10.1186/s12967-023-04132-4

Figure Lengend Snippet: CircEPHA3 inhibits the migration and invasion of PCa cells via miR-513a-3p/BMP2. A Up-regulation of circEPHA3 abolished the cell proliferative capacity of PC-3 and DU145 cells enhanced by miR-513a-3p mimics. B, E The migration capability of PC-3 and DU145 cells transfected with miR-513a-3p mimics was counteracted by circEPHA3 in wound healing assays. Scale bars = 100 μm. C, D The migration and invasion capabilities of PC-3 and DU145 cells transfected with miR-513a-3p mimics was neutralized by circEPHA3 in transwell assays. Scale bars = 100 μm. F The western blot showed that circEPHA3 could reverse the effect of miR-513a-3p on BMP2 protein expression in PC-3 and DU145 cells. G, H The migration capability of circEPHA3-overexpressed PC-3 and DU145 cells co-transfected with miR-513a-3p mimics and/or BMP2 siRNA in wound healing assays. Scale bars = 100 μm. I, J The migration and invasion capabilities of PC-3 and DU145 cells co-transfected with miR-513a-3p mimics and/or BMP2 siRNA in transwell assays. Scale bars = 100 μm. K The western blot showed the level of BMP2 protein in PC-3 and DU145 cells that co-transfected with circEPHA3, miR-513a-3p and/or BMP2 siRNA. *, ** and *** represented P < 0.05, 0.01 and 0.001, respectively

Article Snippet: The primary antibodies against BMP2 (A0231) and β-actin (AC026) were purchased from ABclonal Biotechnology Co., Ltd. (China), and the secondary goat anti-rabbit antibody (BA1055) was procured from Boster Biotechnology Co., Ltd. (China).

Techniques: Migration, Transfection, Western Blot, Expressing

Journal: Journal of Translational Medicine

Article Title: Circular RNA EPHA3 suppresses progression and metastasis in prostate cancer through the miR-513a-3p/BMP2 axis

doi: 10.1186/s12967-023-04132-4

Figure Lengend Snippet: CircEPHA3 prevents tumor growth and metastasis in vivo. A The subcutaneously xenograft tumors formed by circEPHA3-overexpressed or vector PCa cells in BALB/c nude mice. B, C The tumor volume and weight of circEPHA3-overexpressed and vector PCa groups. D The expression of BMP2 protein in circEPHA3-overexpressed and vector PCa groups was assessed by IHC analysis. The red scale bars = 100 μm, the black scar bars = 20 μm. E Representative images of bioluminescence in both metastasis groups captured by IVIS 100 Imaging System. F Representative images of H&E of the lungs in circEPHA3-overexpressed and vector PCa groups. The red scale bars = 500 μm, the black scar bars = 20 μm. *, ** and *** represented P < 0.05, 0.01 and 0.001, respectively

Article Snippet: The primary antibodies against BMP2 (A0231) and β-actin (AC026) were purchased from ABclonal Biotechnology Co., Ltd. (China), and the secondary goat anti-rabbit antibody (BA1055) was procured from Boster Biotechnology Co., Ltd. (China).

Techniques: In Vivo, Plasmid Preparation, Expressing, Imaging

Journal: Journal of Translational Medicine

Article Title: Circular RNA EPHA3 suppresses progression and metastasis in prostate cancer through the miR-513a-3p/BMP2 axis

doi: 10.1186/s12967-023-04132-4

Figure Lengend Snippet: The schematic illustration of circEPHA3 inhibiting PCa cell proliferation and metastasis via the miR-513a-3p/BMP2 axis

Article Snippet: The primary antibodies against BMP2 (A0231) and β-actin (AC026) were purchased from ABclonal Biotechnology Co., Ltd. (China), and the secondary goat anti-rabbit antibody (BA1055) was procured from Boster Biotechnology Co., Ltd. (China).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Angiogenesis-Based Scaffold of MesoporousBioactive Glass Nanofiber on Osteogenesis

doi: 10.3390/ijms232012670

Figure Lengend Snippet: Osteogenic effect of BMSCs cultured on MBG-NH 2 , MBG-NH 2 /IGF and MBG-NH 2 /IGF@SF/VEGF scaffolds in vitro . ( A – D ) Osteogenic mRNA expression levels of BMSCs cultured on the scaffolds for 7 days: Bmp2 ( A ), Opn ( B ), Osterix ( C ), and Runx2 ( D ). ( E ) Runx2 activity of BMSCs transfected with Runx2-binding element-Luc and cultured on the scaffolds for 3 days. Runx2 activity was defined by relative luciferase activity, with renilla luciferase plasmid pRL-SV40 as internal control. ( F ) Osteogenic protein expression levels of BMSCs cultured on the scaffolds for 7 days according to Western blot. Total Erk1/2 and mTOR were used as loading control for p-Erk1/2 and p-mTOR, respectively. ( G – H ) Osteogenic protein expression levels of BMSCs cultured on the scaffolds for 7 days according to immunofluorescence. Actin was used as a loading control for Runx2 ( G ) and Bmp2 ( H ), stained with FITC. Runx2 and Bmp2 (red) were stained with Alexa Fluor 594, and the nuclei (blue) were stained with DAPI. Scale bars: 50 μm. Experimental data were analyzed as indicated and reported as mean ± SE. (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: After being treated with the primary antibody against Bmp2 (Beyotime Biotech, Haimen, China, 1:200), Runx2 (Cell Signaling Technology, USA, 1:1600), CD31(Cell Signaling Technology, Danvers, MA, USA, 1:200) and β-actin (Beyotime Biotech, Haimen, China, 1:20) overnight, β-actin was labeled with FITC (1:100), while Bmp2, Runx2 and CD31 were labeled with Alexa Fluor 594 (1:100).

Techniques: Cell Culture, In Vitro, Expressing, Activity Assay, Transfection, Binding Assay, Luciferase, Plasmid Preparation, Control, Western Blot, Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Angiogenesis-Based Scaffold of MesoporousBioactive Glass Nanofiber on Osteogenesis

doi: 10.3390/ijms232012670

Figure Lengend Snippet: Bone repair ability evaluation of MBG-NH 2 , MBG-NH 2 /IGF and MBG-NH 2 /IGF@SF/VEGF scaffolds in vivo . ( A ) Micro-CT imaging analysis of skull defects in mice after 4 weeks of scaffold transplantation. NC: negative control. ( B ) Tb.N analysis of new bone. ( C ) Histological analysis of skull defects in mice after 4 weeks of scaffold transplantation; HE (top two lines) and Masson’s trichrome (bottom two lines) staining are shown. Scale bars in upper line in HE and Masson’s trichrome staining represent 250 μm; scale bars in the nether line in HE and Masson’s trichrome staining represent 50 μm. NB: native bone; RB: regenerated bone. ( D ) Immunohistochemical analysis of bone defect section after 4 weeks of scaffold transplantation. Bmp2 (top two lines) and CD31 (bottom two lines) staining are shown. Scale bars in upper line in Bmp2 and CD31 staining represent 250 μm; scale bars in the nether line in Bmp2 and CD31 staining represent 50 μm. High expression areas of Bmp2 are indicated by red arrows. Vessels in native bone are indicated by black arrows, and newly formed vessels in regenerated bone are indicated by blue arrows. Experimental data were analyzed as indicated and reported as mean ±SE. (* p < 0.05; ** p < 0.01).

Article Snippet: After being treated with the primary antibody against Bmp2 (Beyotime Biotech, Haimen, China, 1:200), Runx2 (Cell Signaling Technology, USA, 1:1600), CD31(Cell Signaling Technology, Danvers, MA, USA, 1:200) and β-actin (Beyotime Biotech, Haimen, China, 1:20) overnight, β-actin was labeled with FITC (1:100), while Bmp2, Runx2 and CD31 were labeled with Alexa Fluor 594 (1:100).

Techniques: In Vivo, Micro-CT, Imaging, Transplantation Assay, Negative Control, Staining, Immunohistochemical staining, Expressing