Journal: Journal of Natural Medicines

Article Title: Inhibitory functions of cornuside on TGFBIp-mediated septic responses

doi: 10.1007/s11418-021-01601-2

Figure Lengend Snippet: Structures of collismycin C (CNS) and its effects on the release of TGFBIp and expression of receptors. A Structures of CNS. B HUVECs were treated with the indicated concentrations of CNS for 6 h after stimulation with LPS (100 ng/ml, 1 h), and TGFBIp release was measured by ELISA. C Male C57BL/6 mice that underwent CLP were intravenously administered CNS at 0.08–0.8 mg/kg 12 h after CLP ( n = 5). Mice were euthanized 24 h after CLP. Serum TGFBIp levels were measured by ELISA. D The same procedure as in ( B ) except that real-time qRT-PCR analysis was performed using specific primers for TGFBIp and actin, as described in the materials and methods section. E Confluent HUVECs were activated with LPS (100 ng/ml, 3 h), followed by incubation with CNS for 6 h. Expression of αv β3 (white column) and αv β5 (black column) was determined by cell-based ELISA. F Effect of CNS on cellular viability was measured by the MTT assay. D = 0.2% DMSO (vehicle control). Results are expressed as the means ± SD of at least 3 independent experiments ( n = 5/group). * p < 0.05 relative to DMSO ( E left), LPS ( B , D , E right) or CLP ( C ), one-way ANOVA

Article Snippet: The same experimental procedure was used to monitor the cell surface expression of αv β3 and αv β5 using specific antibodies obtained from EMD Millipore (MA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Incubation, In-Cell ELISA, MTT Assay, Control

Journal: Journal of Natural Medicines

Article Title: Inhibitory functions of cornuside on TGFBIp-mediated septic responses

doi: 10.1007/s11418-021-01601-2

Figure Lengend Snippet: Effects of CNS on TGFBIp-mediated proinflammatory responses. A – C , HUVECs were stimulated with TGFBIp (5 μg/ml) for 6 h, followed by treatment with CNS for 6 h. TGFBIp-mediated A expression of VCAM (white column), ICAM (gray column), and E-selectin (black column) in HUVECs, B adherence of human neutrophils to HUVEC monolayers, and C migration of human neutrophils through HUVEC monolayers were analyzed. D The effects of post-treatment with CNS at 0.08–0.8 mg/kg on leukocyte migration into the peritoneal cavities of mice caused by TGFBIp (0.1 mg/kg, i.v.) were analyzed. Results are expressed as the means ± SD of at least 3 independent experiments ( n = 5/group). * p < 0.05 relative to TGFBIp, one-way ANOVA

Article Snippet: The same experimental procedure was used to monitor the cell surface expression of αv β3 and αv β5 using specific antibodies obtained from EMD Millipore (MA).

Techniques: Expressing, Migration

Journal: Molecular Vision

Article Title: Establishment of a continuous untransfected human corneal endothelial cell line and its biocompatibility to denuded amniotic membrane

doi:

Figure Lengend Snippet: Immunofluorescence staining patterns of cell-junction proteins of HCE cells at passage 101. A : human zonula occludens protein 1 (ZO-1). B : human N-cadherin. C : human connecxin-43. D : human integrin αv/β5. Positive expression of the cell-junction proteins of HCE cells was shown. Scale bar: 50 μm.

Article Snippet: Immunofluorescence staining of cell-junction protein expression of HCE cells at passage 101 was performed with goat anti-human zonula occludens protein 1 (ZO-1) polyclonal antibody (1:50; Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-human N-cadherin monoclonal antibody (1:50; Santa Cruz), mouse anti-human connexin 43 monoclonal antibody (1:250; Chemicon, Temecula, CA), mouse anti-human integrin αv/β5 monoclonal antibody (1:50; Santa Cruz), and fluorescent isothiocyanate (FITC)-conjugated rabbit anti-goat IgG antibody or rhodamine B isothiocyanate (RBITC)-conjugated goat anti-mouse IgG antibody or FITC-conjugated goat anti-mouse IgG antibody (1:100; Biosynthesis Biotechnology) as previously described [ ], and visualized with a Nikon Eclipse TE2000-U inverted fluorescent microscope (Nikon, Tokyo, Japan).

Techniques: Immunofluorescence, Staining, Expressing

Journal: Molecular Vision

Article Title: Establishment of a continuous untransfected human corneal endothelial cell line and its biocompatibility to denuded amniotic membrane

doi:

Figure Lengend Snippet: Immunofluorescence staining patterns of cell-junction proteins of the HCE cell sheet formed on dAM. A : human zonula occludens protein 1 (ZO-1). B : human N-cadherin. C : human connexin-43. D : human integrin αv/β5. Positive expression of the cell-junction proteins of passage 101 HCE cells on dAM was shown. Scale bar: 50 μm.

Article Snippet: Immunofluorescence staining of cell-junction protein expression of HCE cells at passage 101 was performed with goat anti-human zonula occludens protein 1 (ZO-1) polyclonal antibody (1:50; Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-human N-cadherin monoclonal antibody (1:50; Santa Cruz), mouse anti-human connexin 43 monoclonal antibody (1:250; Chemicon, Temecula, CA), mouse anti-human integrin αv/β5 monoclonal antibody (1:50; Santa Cruz), and fluorescent isothiocyanate (FITC)-conjugated rabbit anti-goat IgG antibody or rhodamine B isothiocyanate (RBITC)-conjugated goat anti-mouse IgG antibody or FITC-conjugated goat anti-mouse IgG antibody (1:100; Biosynthesis Biotechnology) as previously described [ ], and visualized with a Nikon Eclipse TE2000-U inverted fluorescent microscope (Nikon, Tokyo, Japan).

Techniques: Immunofluorescence, Staining, Expressing