human hbegf elisa kit  (Boster Bio)

Journal: Antioxidants (Basel, Switzerland)

Article Title: Endothelial Notch Signaling Regulates the Function of the Retinal Pigment Epithelial Barrier via EC Angiocrine Signaling.

doi: 10.3390/antiox12111979

Figure Lengend Snippet: Figure 5. Endothelial Notch signaling affects RPE barrier function by regulating the HBEGF paracrine of EC. (A) Analysis for previously published transcriptome sequencing data of normal HUVECs and DAPT treated HUVECs: the GSEA results for ECM (left) and heatmap of the expression of ECM- related genes (right). (B) mRNA and protein levels of HBEGF in normal HUVECs and DAPT-treated HUVECs. (C) mRNA and protein levels of HBEGF in normal HUVECs and NICD adenovirus-infected HUVECs. (D) The secretion levels of HBEGF protein in normal HUVECs and DAPT-treated HUVECs or NICD adenovirus-infected HUVECs were tested using an ELISA kit. (E) Evaluating the RPE barrier function in RPE cells cultured in normal medium, RPE cultured in the EC-conditioned medium, RPE cultured in Notch signaling-inhibited EC-conditioned medium, and RPE cultured in Notch signaling-inhibited EC-conditioned medium supplemented with recombinant HBEGF (r-HBEGF)

Article Snippet: Then, the concentration of secreted HBEGF in the media was measured using the human HBEGF ELISA kit (Boster, Wuhan, China).

Techniques: Sequencing, Expressing, Infection, Enzyme-linked Immunosorbent Assay, Cell Culture, Recombinant

Journal: Antioxidants (Basel, Switzerland)

Article Title: Endothelial Notch Signaling Regulates the Function of the Retinal Pigment Epithelial Barrier via EC Angiocrine Signaling.

doi: 10.3390/antiox12111979

Figure Lengend Snippet: Figure 6. r-HBEGF improved the RPE barrier dysfunction of Notch signaling deletion mice. (A) In the SI-induced RPE barrier disfunction model, whole-mount choroidal staining for EC-specific Notch signaling deletion mice CDH5-CRE-RBPJf/f intravitreally injected with IgG or r-HBEGF, and control mice (RBPJf/+) at p30, were analyzed using ZO-1 staining. The number of RPE cells with intact tight junctions were quantitatively compared. Red circles indicate the location of the optic disk. Scale bars: 50 µm. (B) In the laser-induced CNV model, whole-mount choroidal staining for EC-specific Notch signaling deletion mice CDH5-CRE-RBPJf/f intravitreally injected with IgG or r-HBEGF and control mice (RBPJf/+) were analyzed using IB-4 staining. IB4 positive areas were quantitatively compared. Three biological replicates were performed and values are presented as the mean ± SEM (n = 3). Scale bars: 150 µm. * p < 0.05, ** p < 0.01.

Article Snippet: Then, the concentration of secreted HBEGF in the media was measured using the human HBEGF ELISA kit (Boster, Wuhan, China).

Techniques: Staining, Injection, Control

Journal: Antioxidants (Basel, Switzerland)

Article Title: Endothelial Notch Signaling Regulates the Function of the Retinal Pigment Epithelial Barrier via EC Angiocrine Signaling.

doi: 10.3390/antiox12111979

Figure Lengend Snippet: Figure 7. EC regulates MMP-9 expression in RPE cells via Endothelial Notch signaling-driven HBEGF secretion. (A) Protein interaction prediction analysis results of HBEGF and MMP-9, using the String database. (B) mRNA level, protein level, and enzyme activity level of MMP-9 in the RPE, RPE with HUVEC, and RPE with Notch signaling-inhibited HUVECs were tested via qRT-PCR, Western blot, and gelatin assay kit. (C) mRNA levels of MMP-9 in normal HUVECs and DAPT-treated HUVECs were tested via qRT-PCR. (D,E) mRNA and protein levels of MMP-9 in the RPE, RPE with HUVECs, RPE with siNC-treated HUVECs, and RPE with siHBEGF-treated HUVECs were tested via qRT-PCR and Western blot. (F) Protein levels of MMP-9 in RPE cells cultured in normal medium, RPE cultured in EC-conditioned medium, RPE cultured in Notch signaling-inhibited EC-conditioned medium, and RPE cultured in Notch signaling-inhibited EC-conditioned medium supplemented with recombinant HBEGF (r-HBEGF) were tested using Western blot. Three biological replicates were performed and values are presented as the mean ± SEM (n = 3). NS, no significance, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Then, the concentration of secreted HBEGF in the media was measured using the human HBEGF ELISA kit (Boster, Wuhan, China).

Techniques: Expressing, Activity Assay, Quantitative RT-PCR, Western Blot, Cell Culture, Recombinant

Journal: Oncogene

Article Title: Increased metabolites of 5-lipoxygenase from hypoxic ovarian cancer cells promote tumor-associated macrophage infiltration.

doi: 10.1038/onc.2014.85

Figure Lengend Snippet: Figure 2. Key proteins and metabolites of the 5-LOX pathway are increased in hypoxic ovarian cancer cells. (a) SKOV-3 and OVCAR-3 ovarian cancer cells were treated with 1% O2 for 0, 4, 8, 12 and 24 h. Transcript levels of 5-LOX, FLAP and LTA4H were analyzed by quantitative reverse transcriptase-PCR. *Po0.05 versus 0 h. (b) SKOV-3 cells were treated with 1% O2 for 0, 6, 12, 24 and 48 h. The expression of HIF-1α, 5-LOX and FLAP were analyzed by western blotting. *Po0.05 versus Con. (c) SKOV-3 cells were treated with 1% O2 for 6, 12, 24, 48 and 72 h. CM was harvested and analyzed by ELISA. *Po0.05 versus normoxia. (d) SKOV-3 cells were transfected with siRNAs and treated with 1% O2 for 24 h. The expression of HIF-1α, 5-LOX and FLAP were analyzed by western blotting. *Po0.05 versus Con siRNA. #Po0.05 versus corresponding Con siRNA.

Article Snippet: For measurement of secreted HB-EGF and TNF-α, levels of HB-EGF and TNF-α present in the supernatants were measured using the human HB-EGF ELISA kit (Cusabio Biotech Co., Ltd) and human TNF-α ELISA Kit (R&D Systems) following the manufacture’s protocol.

Techniques: Reverse Transcription, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Transfection

Journal: Oncogene

Article Title: Increased metabolites of 5-lipoxygenase from hypoxic ovarian cancer cells promote tumor-associated macrophage infiltration.

doi: 10.1038/onc.2014.85

Figure Lengend Snippet: Figure 6. Metabolites of 5-LOX enhance release of TNF-α and HB-EGF through upregulation of MMP-7. TNF-α and HB-EGF in medium were detected by ELISA. *Po0.05 versus Con. #Po0.05 versus corresponding CM, CM+isotype-IgG or isotype-IgG. Macrophages were cultured with medium alone (Con), normoxia (N) or hypoxia (H) CM from SKOV-3 cells (a) or untreated and MK886-treated SKOV-3 cells (b). (c) Isotype-IgG or anti-MMP-7 were added in the CM. Macrophages were treated with 5-HETE (d) or LTB4 (e). (f) Macrophages were treated with 5-HETE or LTB4. Then isotype-IgG or anti-MMP-7 were added in the medium.

Article Snippet: For measurement of secreted HB-EGF and TNF-α, levels of HB-EGF and TNF-α present in the supernatants were measured using the human HB-EGF ELISA kit (Cusabio Biotech Co., Ltd) and human TNF-α ELISA Kit (R&D Systems) following the manufacture’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture