Journal: Bone research

Article Title: Metformin accelerates bone fracture healing by promoting type H vessel formation through inhibition of YAP1/TAZ expression.

doi: 10.1038/s41413-023-00279-4

Figure Lengend Snippet: Fig. 1 Metformin promotes angiogenesis under hypoxic conditions in vitro. Representative images (a) of the transwell migration assay with quantification of crystal violet-stained migrated HMECs (b) treated with metformin under different oxygen concentrations (21% and 1% O2). Met: metformin. Scale bar: 100 μm. n = 3 per group. Representative images (c) of tube formation of HMECs on Matrigel with quantification of the total branching points (d), total tube length (e), and total loop numbers (f). Scale bar: 200 μm. n = 3 per group. g CCK-8 analysis of the proliferation of HMECs. n = 4 per group. h qRT‒PCR analysis of the mRNA levels of the HIF-1α target genes Vegfa and Lrg1 in HMECs with or without metformin treatment under different oxygen concentrations. n = 3 per group. Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) The concentrations of OCN and VEGFA were determined using the Mouse OCN (Elabscience, E-EL-M0864c, Wuhan, China) or VEGFA (Elabscience, E-EL-M1292c) ELISA kit according to the manufacturers’ instructions.

Techniques: In Vitro, Transwell Migration Assay, Staining, CCK-8 Assay

Journal: Bone research

Article Title: Metformin accelerates bone fracture healing by promoting type H vessel formation through inhibition of YAP1/TAZ expression.

doi: 10.1038/s41413-023-00279-4

Figure Lengend Snippet: Fig. 3 Metformin promotes type H vessel formation in osteoporotic fracture mice. a Representative CD31 and Emcn coimmunostaining images (left) with quantification of the type H vessel ratio in calluses (right) from the osteoporotic mice treated with PBS, Met, PTH, and ALN at 3, 6, and 9 weeks post-fracture. ca: callus. The dotted line represents the boundary of the callus. Met metformin, ALN alendronate, PTH parathyroid hormone. Scale bar: 100 μm. n = 5 per group. b Representative Ki67 and Emcn coimmunostaining images (left) with quantification of the number of Ki67-positive endothelial cells in calluses (right) from the osteoporotic mice treated with PBS, Met, PTH, and ALN at 3, 6, and 9 weeks post-fracture. Scale bar: 100 μm. n = 5 per group. ELISAs for the serum (c) and bone marrow (d) concentrations of VEGFA at 9 weeks post-osteoporotic fracture. n = 8 per group; Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) The concentrations of OCN and VEGFA were determined using the Mouse OCN (Elabscience, E-EL-M0864c, Wuhan, China) or VEGFA (Elabscience, E-EL-M1292c) ELISA kit according to the manufacturers’ instructions.

Techniques:

Journal: Bone research

Article Title: Metformin accelerates bone fracture healing by promoting type H vessel formation through inhibition of YAP1/TAZ expression.

doi: 10.1038/s41413-023-00279-4

Figure Lengend Snippet: Fig. 4 Metformin promotes the expression of HIF-1α by inhibiting the expression of YAP1/TAZ in HMECs under hypoxic conditions. a qRT‒PCR analysis of the inhibitory efficiency of siRNAs targeting HIF-1α. n = 3 per group. b qRT‒PCR analysis of the expression of HIF-1α and its target genes Vegfa and Lrg1 in the si-HIF-1α-transfected HMECs with or without metformin treatment under hypoxic conditions (1% O2). Met: metformin. n = 3 per group. Immunofluorescence staining images and quantification showing the protein levels of HIF-1α (c), YAP1 (d), and TAZ (e) in the HMECs treated with PBS (control) or metformin under hypoxic conditions. Scale bar: 20 μm. n = 9 per group. f qRT‒PCR analysis of the inhibitory efficiency of siRNAs targeting YAP1 or TAZ. n = 3 per group. g Immunofluorescence staining images and quantification showing the protein level of HIF-1α in the hypoxia-cultured HMECs from the si-Con, si-YAP1, si-TAZ, si-Y/T, and si-Y/T + Met groups. Y/T: YAP1 and TAZ. Scale bar: 20 μm. n = 3 per group. Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) The concentrations of OCN and VEGFA were determined using the Mouse OCN (Elabscience, E-EL-M0864c, Wuhan, China) or VEGFA (Elabscience, E-EL-M1292c) ELISA kit according to the manufacturers’ instructions.

Techniques: Expressing, Transfection, Staining, Control, Cell Culture

Journal: Chinese Journal of Cancer Research

Article Title: Tim-3 promotes cell aggressiveness and paclitaxel resistance through NF-κB/STAT3 signalling pathway in breast cancer cells

doi: 10.21147/j.issn.1000-9604.2020.05.02

Figure Lengend Snippet: Effect of Tim-3 OE on tube formation of endothelial cells. (A) Tube formation ability of HUVECs cultured in conditioned medium from MDA-MB-231 Tim-3-overexpressing cells (P=0.014 vs . Scr); (B) Tube formation ability of HUVECs cultured in conditioned medium from MCF7 Tim-3-overexpressing cells (P=0.016 vs . Scr); (C) Protein levels of VEGFA, VEGFB and VEGFD following Tim-3 OE (left) and quantitative densitometric analysis (right); (D) mRNA expression of VEGFA and VEGFD genes in breast cancer cells. OE, overexpression; Tim-3, T-cell immunoglobulin and mucin-domain containing molecule-3; HUVEC, human umbilical vein endothelial cell; VEGF, vascular endothelial growth factor. * , P<0.05; ** , P<0.01; *** , P<0.001.

Article Snippet: ELISA was performed using the human VEGFC (cat. no. E-EL-H1600; Elabscience) and vascular endothelial growth factor receptor 2 (VEGFR2) ELISA kit (cat. no. E-EL-H1603; Elabscience), according to the manufacturer’s protocol.

Techniques: Cell Culture, Expressing, Over Expression

Journal: Chinese Journal of Cancer Research

Article Title: Tim-3 promotes cell aggressiveness and paclitaxel resistance through NF-κB/STAT3 signalling pathway in breast cancer cells

doi: 10.21147/j.issn.1000-9604.2020.05.02

Figure Lengend Snippet: VEGFC and VEGFR2 protein levels in conditioned media of stable cell lines indicated by ELISA. (A) VEGFC; (B) VEGFR2. VEGF, vascular endothelial growth factor; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: ELISA was performed using the human VEGFC (cat. no. E-EL-H1600; Elabscience) and vascular endothelial growth factor receptor 2 (VEGFR2) ELISA kit (cat. no. E-EL-H1603; Elabscience), according to the manufacturer’s protocol.

Techniques: Stable Transfection, Enzyme-linked Immunosorbent Assay

Journal: Chinese Journal of Cancer Research

Article Title: Tim-3 promotes cell aggressiveness and paclitaxel resistance through NF-κB/STAT3 signalling pathway in breast cancer cells

doi: 10.21147/j.issn.1000-9604.2020.05.02

Figure Lengend Snippet: Schematic illustration of role of Tim-3 in breast cancer. Upregulation of Tim-3 not only promotes cell proliferation, migration and invasion, but also disrupts cell-cell tight junction, increases angiogenesis of endothelial cells and paclitaxel-resistance. Tim-3 functions in breast cancer cells by activating NF-κB/STAT3 pathway and downstream target genes. Tim-3, T-cell immunoglobulin and mucin-domain containing molecule-3; IL-6, interleukin 6; ZO, zona occludens; VEGF, vascular endothelial growth factor; CCND1, cyclin D1; MMP-1, matrix metalloproteinase-1.

Article Snippet: ELISA was performed using the human VEGFC (cat. no. E-EL-H1600; Elabscience) and vascular endothelial growth factor receptor 2 (VEGFR2) ELISA kit (cat. no. E-EL-H1603; Elabscience), according to the manufacturer’s protocol.

Techniques: Migration

Journal: Experimental and therapeutic medicine

Article Title: Expression and predictive value of HIF-1α and VEGF in patients with burns following treatment.

doi: 10.3892/etm.2020.9270

Figure Lengend Snippet: Figure 1. Changes in serum HIF‑1α and VEGF levels before and after treatment. (A) HIF‑1α decreased significantly following treatment (t=4.983, P<0.001). (B) VEGF significantly increased following treatment (t=8.826, P<0.001). ***P<0.001 vs. before treatment. HIF, hypoxia‑inducible factor; VEGF, vascular endothelial growth factor.

Article Snippet: HIF‐1α ELISA detection kit (cat. no. E‐EL‐H6066) and VEGF ELISA assay kit (cat. no. E‐EL‐H1601c) were purchased from Elabscience Biotechnology Co., Ltd. Moisturizing Burn Cream was obtained from Mebo Pharmaceutical Co., Ltd.

Techniques:

Journal: Experimental and therapeutic medicine

Article Title: Expression and predictive value of HIF-1α and VEGF in patients with burns following treatment.

doi: 10.3892/etm.2020.9270

Figure Lengend Snippet: Figure 2. Predictive value of HIF‑1α and VEGF for treatment efficacy. (A) The level of HIF‑1α in the ineffective group was significantly higher compared with the effective group (t=3.767, P<0.001). (B) VEGF levels in the ineffective group was significantly lower compared with the effective group (t=4.542, P<0.001). ***P<0.001. (C) The AUC of HIF‑1α for treatment efficacy was 0.795, and when the cut‑off point was 161.757, its optimal specificity and sensitivity were 68.75 and 80.88%, and the Youden index was 49.63%. The AUC of VEGF for treatment efficacy was 0.826, and when the cut‑off point was 437.406, the optimal specificity and sensitivity were 68.75 and 82.35% respectively, and the Youden index was 51.10%. While the AUC of the joint detection for treatment efficacy was 0.847, and when the cut‑off point was set as 0.847, the optimal specificity and sensitivity were 87.50 and 66.18% and the Youden index was 53.68%. HIF, hypoxia‑inducible factor; VEGF, vascular endothelial growth factor; AUC, area under the curve.

Article Snippet: HIF‐1α ELISA detection kit (cat. no. E‐EL‐H6066) and VEGF ELISA assay kit (cat. no. E‐EL‐H1601c) were purchased from Elabscience Biotechnology Co., Ltd. Moisturizing Burn Cream was obtained from Mebo Pharmaceutical Co., Ltd.

Techniques: