Journal: Journal of Cellular and Molecular Medicine

Article Title: Apatinib inhibits the growth of small cell lung cancer via a mechanism mediated by VEGF, PI3K/Akt and Ki‐67/CD31

doi: 10.1111/jcmm.16926

Figure Lengend Snippet: Levels of VEGF, VEGFR2 and pVEGFR2 in NCI‐H345 and NCI‐H446 cells which were examined by Western blotting. All blots were normalized to GAPDH levels. * p < 0.05 vs. NCI‐H345 cells. VEGF, vascular endothelial growth factor; VEGFR2, vascular endothelial growth factor receptor 3; p, phosphorylated

Article Snippet: The levels of VEGF, VEGFR2, and pVEGFR2 in NCI‐H345 and NCI‐H446 cells were examined by Western blot analysis to select the ideal cell model. NCI‐H345 and NCI‐H446 cells were subsequently treated with 30 ng/ml recombinant human VEGF (rhVEGF; cat. no 10542‐H08H; Sino Biological) for 0, 30, 60 and 120 min at 37°C.

Techniques: Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Apatinib inhibits the growth of small cell lung cancer via a mechanism mediated by VEGF, PI3K/Akt and Ki‐67/CD31

doi: 10.1111/jcmm.16926

Figure Lengend Snippet: Levels of VEGFR2 and pVEGFR2 in NCI‐H345 cells following treatment with treatment with rhVEGF for 0, 30, 60 and 120 min analysed by Western blotting. All blots were normalized to GAPDH levels. * p < 0.05 vs. control. VEGFR2, vascular endothelial growth factor receptor 3; p, phosphorylated; rhVEGF, recombinant human VEGF

Article Snippet: The levels of VEGF, VEGFR2, and pVEGFR2 in NCI‐H345 and NCI‐H446 cells were examined by Western blot analysis to select the ideal cell model. NCI‐H345 and NCI‐H446 cells were subsequently treated with 30 ng/ml recombinant human VEGF (rhVEGF; cat. no 10542‐H08H; Sino Biological) for 0, 30, 60 and 120 min at 37°C.

Techniques: Western Blot, Recombinant

Journal: Journal of Cellular and Molecular Medicine

Article Title: Apatinib inhibits the growth of small cell lung cancer via a mechanism mediated by VEGF, PI3K/Akt and Ki‐67/CD31

doi: 10.1111/jcmm.16926

Figure Lengend Snippet: The proliferation rate of NCI‐H345 cells (A) following treatments with increasing concentrations of apatinib (50, 100, 150, 200 and 250 nM) for different durations (24, 48 and 72 h) and (B) after various treatments of rhVEGF, apatinib and serum‐free medium for 60 min, as evaluated by Cell Counting Kit‐8 assay. * p < 0.05 vs. control; # p < 0.05 vs. SM+apatinib. SM, serum‐free medium

Article Snippet: The levels of VEGF, VEGFR2, and pVEGFR2 in NCI‐H345 and NCI‐H446 cells were examined by Western blot analysis to select the ideal cell model. NCI‐H345 and NCI‐H446 cells were subsequently treated with 30 ng/ml recombinant human VEGF (rhVEGF; cat. no 10542‐H08H; Sino Biological) for 0, 30, 60 and 120 min at 37°C.

Techniques: Cell Counting

Journal: Oncogene

Article Title: Neuropilin-2 regulates androgen-receptor transcriptional activity in advanced prostate cancer

doi: 10.1038/s41388-022-02382-y

Figure Lengend Snippet: Following ectopic expression of untagged NRP2B in C4–2B cells, nuclear and post-nuclear fractions were separated, and a pull-down assay was performed on the nuclear fraction with NRP2 antibody. Mass-Spectrometry was carried out with the pull-down samples. Using the genes detected in NRP2 Mass-Spectrometry. A. Sum of all the spectra associated with NRP2B pulldown sample is determined by the spectral graph. B. Venn-diagram represent the overlapping genes identified in two independent NRP2B mass-spectrometry assay. C. Group of nuclear pore proteins (Nups) identified through NRP2B Mass-Spectrometry. Schematic diagram indicating the relative positions of Nups around the nuclear pore. D. Validation of NRP2-Nup93 interactions in C4–2B. C4–2B cells were transfected with HA-tagged NRP2B. IP was carried out with HA-antibody and immunoblots were carried out with anti-Nup93. E. Endogenously, NRP2 and Nup93 interaction was validated in C4–2B and 22Rv1 cell lines. NRP2 was pull down with NRP2-specific antibody and IB with Nup93 antibody. F. NRP2B and AR interaction was carried out in C4–2B following ectopic interaction of HA-tagged NRP2B. Pulldown was carried out with HA-antibody and IP was carried out against NRP2 and AR. G. NRP2 and AR interaction was also monitored in PC3 and 22Rv1 following ectopic expression of HA-tagged NRP2B. IB was carried out against HA and AR. H. Co-IP for AR with NRP2 was carried out with pulldown of NRP2 by HA-antibody in C4–2B cells expressing wild type and mutant K892A NRP2B isoforms. NRP2B immunoblot was carried out with HA-antibody. I. IP with AR antibody was carried out to test the interaction among AR, Nup93 and NRP2 under the presence or absence of NRP2 and Nup 93 from the nuclear fraction of LNCaP C4–2B cells. An immunoblot was performed with anti-AR, anti-Nup93 and anti-NRP2 antibody. The Co-IP was carried out in the presence of 50ng/ml VEGF-C. J. PLA was carried out to validating the NRP2-AR interaction within the nucleus. Following ectopic expression of HA-tagged NRP2B, PLA was carried out with HA and endogenous AR antibodies under the presence or absence of NUP93. As a negative control, only HA-antibody was used for PLA reaction. Arrowhead indicates the immune-reactive PLA puncta. Nucleus was counter-stained with NUP98 to demarcate the nuclear periphery. DAPI used for nuclear staining. PLA quantification was shown in Bar diagram.

Article Snippet: NRP2B or NRP2A localization were chased with various experimental treatment condition for 1hrs under the following reagents VEGF-C (50ng/ml, R&D System, 752-VC-025), NRP2Fc (100ng/ml, R&D System, 2215-N2–025), SEMA3F (100ng/ml, R&D System, 9878-S3–025), Brefeldin A (0.5μM, Sigma, B6542).

Techniques: Expressing, Pull Down Assay, Mass Spectrometry, Biomarker Discovery, Transfection, Western Blot, Co-Immunoprecipitation Assay, Mutagenesis, Negative Control, Staining

Journal: Communications Biology

Article Title: Preventing periprosthetic osteolysis in aging populations through lymphatic activation and stem cell-associated secretory phenotype inhibition

doi: 10.1038/s42003-024-06664-x

Figure Lengend Snippet: a Micro-CT images illustrating cranial bone resorption from both superior and inferior perspectives after continuous TAP treatment for 7 or 14 days, concomitant with subcutaneous injection of recombinant VEGF-C. b TRAP staining in mouse cranial bones following TAP implantation, combined with recombinant VEGF-C injection. Scale bars: 100 μm. c H&E staining in mouse cranial bones following TAP implantation, combined with recombinant VEGF-C injection. Scale bars: 100 μm. d IF staining for DAPI with LYVE1 (green) and PROX1 (red). Scale bars, 100 μm. e Quantification of the lytic area in calvarial bone tissues analyzed by micro-CT ( n = 6). f Quantification of TRAP-stained areas in calvarial bone sections ( n = 6). g Quantification of inflammatory infiltrating cells in calvarial bone sections ( n = 6). h , i The corresponding quantitative data for LYVE1 (green) and PROX1 (red) expressions in cranial bones on day 7 or day 14 after combined TAP and recombinant VEGF-C treatments ( n = 6). Data are shown as means ± SD. Significance was assessed through one-way ANOVA.

Article Snippet: For pharmacological activation of lymphatic vessels in mouse bone tissue, recombinant VEGF-C protein (SinoBiological) at a concentration of 10 μg/mL was subcutaneously injected onto the surface of the skull.

Techniques: Micro-CT, Injection, Recombinant, Staining

Journal: Communications Biology

Article Title: Preventing periprosthetic osteolysis in aging populations through lymphatic activation and stem cell-associated secretory phenotype inhibition

doi: 10.1038/s42003-024-06664-x

Figure Lengend Snippet: a Micro-CT images depict cranial bone resorption from both superior and inferior perspectives after continuous LPS treatment for 14 days, concomitant with subcutaneous injection of recombinant VEGF-C. b TRAP staining in mouse cranial bones following LPS treatment, combined with recombinant VEGF-C injection. Scale bars: 100 μm. c H&E staining in mouse cranial bones following LPS treatment, combined with recombinant VEGF-C injection. Scale bars: 100 μm. d IF staining for DAPI with LYVE1 (green) and PROX1 (red). Scale bars, 100 μm. e Quantification of the lytic area in calvarial bone tissues analyzed by micro-CT ( n = 6). f Quantification of TRAP-stained areas in calvarial bone sections ( n = 6). g Quantification of inflammatory infiltrating cells in calvarial bone sections ( n = 6). h , i The corresponding quantitative data for LYVE1 (green) and PROX1 (red) expressions in cranial bones on day 14 after combined LPS and recombinant VEGF-C treatments ( n = 6). j Micro-CT images depict cranial bone resorption from both superior and inferior perspectives after continuous TNF-α treatment for 14 days, concomitant with subcutaneous injection of recombinant VEGF-C. k TRAP staining in mouse cranial bones following TNF-α treatment, combined with recombinant VEGF-C injection. Scale bars: 100 μm. l H&E staining in mouse cranial bones following TNF-α treatment, combined with recombinant VEGF-C injection. Scale bars: 100 μm. m IF staining for DAPI with LYVE1 (green) and PROX1 (red). Scale bars, 100 μm. n Quantification of the lytic area in calvarial bone tissues analyzed by micro-CT ( n = 6). o Quantification of TRAP-stained areas in calvarial bone sections ( n = 6). p Quantification of inflammatory infiltrating cells in calvarial bone sections ( n = 6). q , r The corresponding quantitative data for LYVE1 (green) and PROX1 (red) expressions in cranial bones on day 14 after combined TNF-α and recombinant VEGF-C treatments ( n = 6). Data are shown as means ± SD. Significance was assessed through one-way ANOVA.

Article Snippet: For pharmacological activation of lymphatic vessels in mouse bone tissue, recombinant VEGF-C protein (SinoBiological) at a concentration of 10 μg/mL was subcutaneously injected onto the surface of the skull.

Techniques: Micro-CT, Injection, Recombinant, Staining

Journal: Communications Biology

Article Title: Preventing periprosthetic osteolysis in aging populations through lymphatic activation and stem cell-associated secretory phenotype inhibition

doi: 10.1038/s42003-024-06664-x

Figure Lengend Snippet: a Micro-CT images illustrating cranial bone resorption in aging mice from both superior and inferior perspectives after continuous TAP treatment for 14 days, concomitant with subcutaneous injection of recombinant VEGF-C. b Quantification of the lytic area in calvarial bone tissues analyzed by micro-CT ( n = 6). c TRAP staining in aging mouse cranial bones after TAP implantation. Scale bars: 100 μm. d H&E staining in aging mouse cranial bones after TAP implantation. Scale bars: 100 μm. e IF staining for DAPI with LYVE1 (green) and PROX1 (red). Scale bars, 100 μm. f Quantification of TRAP-stained areas in calvarial bone sections ( n = 6). g Quantification of inflammatory infiltrating cells in calvarial bone sections ( n = 6). h , i The corresponding quantitative data for LYVE1 (green) and PROX1 (red) expressions in the cranial bones of aging mice on day 14 after combined TAP and recombinant VEGF-C treatments ( n = 6). j – l Representative immunofluorescence images and the corresponding quantification data for LYVE1 (green) and PROX1 (red) expressions in the cranial bones of mice at different ages ( n = 6). m – p Representative immunofluorescence images and the corresponding quantification data for p53, γH2AX and Perilipin expressions in the cranial bones of mice at different ages ( n = 6). Data are shown as means ± SD. Significance was assessed through one-way ANOVA.

Article Snippet: For pharmacological activation of lymphatic vessels in mouse bone tissue, recombinant VEGF-C protein (SinoBiological) at a concentration of 10 μg/mL was subcutaneously injected onto the surface of the skull.

Techniques: Micro-CT, Injection, Recombinant, Staining, Immunofluorescence

Journal: Communications Biology

Article Title: Preventing periprosthetic osteolysis in aging populations through lymphatic activation and stem cell-associated secretory phenotype inhibition

doi: 10.1038/s42003-024-06664-x

Figure Lengend Snippet: a Model of co-culture involving BMSCs treated with adipogenic differentiation medium and LECs. Mouse bone marrow-derived stem cells were seeded in the upper insert at a density of 5 × 10 4 cells/well, while the lower layer of LECs was evenly plated at a density of 1 × 10 5 cells/well. b IF images of Anti-LYVE1 antibody and Anti-PROX1 antibody demonstrate that upper-layer adipogenically differentiated BMSCs inhibit the proliferation of lower-layer LECs. However, this inhibitory effect can be reversed by using JAK inhibitor on the upper insert. Scale bar, 50 µm. c Micro-CT images illustrate cranial bone resorption from both superior and inferior perspectives after continuous TAP treatment for 14 days, along with subcutaneous injection of recombinant VEGF-C, oral administration of JAK inhibitor, or their combination. d TRAP staining was performed on mouse cranial bones following TAP treatment, combined with subcutaneous injection of recombinant VEGF-C, oral administration of JAK inhibitor, or their combination. Scale bars: 100 μm. e H&E staining was performed on mouse cranial bones following TAP treatment, combined with subcutaneous injection of recombinant VEGF-C, oral administration of JAK inhibitor, or their combination. Scale bars: 100 μm. f IF staining for DAPI with LYVE1 (green) and PROX1 (red). Scale bars, 100 μm. g Micro-CT images illustrating cranial bone resorption in aging mice from both superior and inferior perspectives following TAP treatment, combined with subcutaneous injection of recombinant VEGF-C, oral administration of JAK inhibitor, or their combination. h Quantification of TRAP-stained areas in calvarial bone sections ( n = 6). i Quantification of inflammatory infiltrating cells in calvarial bone sections ( n = 6). j , k The corresponding quantitative data for LYVE1 (green) and PROX1 (red) expressions in cranial bones on day 14 after combined TAP treatment, subcutaneous injection of recombinant VEGF-C, oral administration of JAK inhibitor, or their combination ( n = 6). Data are shown as means ± SD. Significance was assessed through one-way ANOVA.

Article Snippet: For pharmacological activation of lymphatic vessels in mouse bone tissue, recombinant VEGF-C protein (SinoBiological) at a concentration of 10 μg/mL was subcutaneously injected onto the surface of the skull.

Techniques: Co-Culture Assay, Derivative Assay, Micro-CT, Injection, Recombinant, Staining

Journal: World Journal of Gastroenterology

Article Title: Role of VEGF and CD44v6 in differentiating benign from malignant ascites

doi: 10.3748/wjg.v9.i11.2596

Figure Lengend Snippet: Comparison of VEGF concentrations in different kinds of ascites. Group 1: cirrhotic ascites, Group 2: tuberculous ascites, Group 3: malignant ascites.

Article Snippet: Immunoassay for human VEGF Concentrations of VEGF in ascites were determined with an ELISA kit (R&D Systems) following the manufacturer’s guidelines.

Techniques: Comparison