antibodies against efna5  (Abmart Inc)

Journal: Antioxidants

Article Title: EFNA5 as an Oocyte-Derived Factor Enhances Developmental Competence by Modulating Oxidative Stress, Inflammation, and Apoptosis During In Vitro Maturation

doi: 10.3390/antiox14121476

Figure Lengend Snippet: The expression pattern of EFNA5 and EPHA4 in COCs. ( A ) Cross-species heatmaps of EFNA family expression in GV and MII oocytes (row-scaled Z-scores). ( B ) The expression of EFNA5 transcript in GV and MII oocytes from multiple species. ( C ) Western blots of EFNA5 in cumulus cells and oocytes with densitometry normalized to GAPDH. ( D ) Immunofluorescence localization of EFNA5 in COCs (DAPI, blue; EFNA5, red). Scale bars = 50 µm. ( E ) qRT-PCR quantification of EFNA5 and EphA receptors in oocytes and cumulus cells (row-scaled Z-scores). ( F ) Immunofluorescence of EPHA4 in COCs (DAPI, blue; EPHA4, red). Scale bars = 50 µm. Abbreviations: OO, oocyte; CC, cumulus cell; GV, germinal vesicle; MII, metaphase II. All experiments were repeated at least three times. Data are presented as mean ± SEM.

Article Snippet: Samples were fixed in 4% paraformaldehyde (PFA) at 4 °C for 30 min, permeabilized with 0.1% Triton X-100 in PBS for 15 min at room temperature, and blocked with 1% BSA in PBS for 1 h. Cells were incubated overnight at 4 °C with primary antibodies against EFNA5 (1:100; ABMART, TP72055, Shanghai, China) and EPHA4 (1:100; Proteintech, 21875-1-AP, Wuhan, China).

Techniques: Expressing, Western Blot, Immunofluorescence, Quantitative RT-PCR

Journal: Antioxidants

Article Title: EFNA5 as an Oocyte-Derived Factor Enhances Developmental Competence by Modulating Oxidative Stress, Inflammation, and Apoptosis During In Vitro Maturation

doi: 10.3390/antiox14121476

Figure Lengend Snippet: EFNA5 enhances the oocyte developmental competence. ( A ) Cross-species heatmaps comparing EFNA5 expression IVO and IVM oocytes from human, mouse, sheep and mouse (row-scaled Z-scores). ( B ) Representative bright-field images of embryos derived from oocytes matured with 0, 10, 50, or 100 ng/mL recombinant EFNA5. Scale bar = 200 µm. ( C , D ) Cleavage rate and blastocyst rate for each group. ( E ) Representative images of blastocysts stained with DAPI (blue) and TUNEL (green). Scale bar = 50 µm. ( F , G ) Total cell number and percentage of TUNEL-positive nuclei per blastocyst. Abbreviations: IVO, in vivo maturation; IVM, in vitro maturation. All experiments were repeated at least three times. Data are mean ± SEM. Different letters indicate p < 0.05.

Article Snippet: Samples were fixed in 4% paraformaldehyde (PFA) at 4 °C for 30 min, permeabilized with 0.1% Triton X-100 in PBS for 15 min at room temperature, and blocked with 1% BSA in PBS for 1 h. Cells were incubated overnight at 4 °C with primary antibodies against EFNA5 (1:100; ABMART, TP72055, Shanghai, China) and EPHA4 (1:100; Proteintech, 21875-1-AP, Wuhan, China).

Techniques: Expressing, Derivative Assay, Recombinant, Staining, TUNEL Assay, In Vivo, In Vitro

Journal: Antioxidants

Article Title: EFNA5 as an Oocyte-Derived Factor Enhances Developmental Competence by Modulating Oxidative Stress, Inflammation, and Apoptosis During In Vitro Maturation

doi: 10.3390/antiox14121476

Figure Lengend Snippet: EFNA5 promotes cumulus expansion and suppresses apoptosis in cumulus–oocyte complexes. ( A ) Bright-field images of GV and MII stage COCs matured with 0, 10, 50, or 100 ng/mL EFNA5. Scale bar = 200 µm. ( B ) Distribution of cumulus expansion scores (Score 1, minimal; Score 2, moderate; Score 3, full) under each EFNA5 dose. ( C ) qRT-PCR analysis of cumulus-expansion genes ( PTX3 , HAS2 , PTGS2 , TNFAIP6 ) in COCs after IVM. ( D ) Representative images of TUNEL staining in COCs matured with the indicated EFNA5 doses (DAPI, blue; TUNEL, green). Scale bar = 50 µm. ( E ) Quantification of TUNEL fluorescence intensity in COCs. ( F ) qRT-PCR analysis of apoptosis-related genes ( BCL2 and TP53 ) in COCs. All experiments were repeated at least three times. Data are mean ± SEM. Different letters indicate p < 0.05.

Article Snippet: Samples were fixed in 4% paraformaldehyde (PFA) at 4 °C for 30 min, permeabilized with 0.1% Triton X-100 in PBS for 15 min at room temperature, and blocked with 1% BSA in PBS for 1 h. Cells were incubated overnight at 4 °C with primary antibodies against EFNA5 (1:100; ABMART, TP72055, Shanghai, China) and EPHA4 (1:100; Proteintech, 21875-1-AP, Wuhan, China).

Techniques: Quantitative RT-PCR, TUNEL Assay, Staining, Fluorescence

Journal: Antioxidants

Article Title: EFNA5 as an Oocyte-Derived Factor Enhances Developmental Competence by Modulating Oxidative Stress, Inflammation, and Apoptosis During In Vitro Maturation

doi: 10.3390/antiox14121476

Figure Lengend Snippet: Effects of EFNA5 on the transcriptional profile of oocytes matured in vitro. ( A ) PCA of MII oocyte transcriptomes in control and EFNA5 groups. ( B ) Volcano plot of DEGs; dashed lines indicate |log 2 FC| = 1.5 and p -value = 0.05. Red, upregulated; blue, downregulated; grey, not significant. ( C ) Heatmap of DEGs across samples with hierarchical clustering (row-scaled Z-scores). ( D ) GO enrichment (BP/CC/MF); bar length denotes-log 10 (P), numbers indicate gene counts. Highlighted pathways are mainly associated with inflammation and antioxidant defense. ( E ) KEGG pathway enrichment; bubble size represents gene count, color indicates-log 10 (P), x -axis shows Gene Ratio. Highlighted pathways are mainly associated with inflammation and antioxidant defense. ( F – H ) Module heatmaps for antioxidant (NRF2 targets), inflammatory (NF-κB axis), and apoptotic genes (row-scaled Z-scores). ( I ) GSEA enrichment plots for Glutathione metabolism, NF-κB signaling, and p53 pathway. ( J ) PPI network of core DEGs with hub nodes (node size = degree; edge thickness = interaction confidence). Abbreviations: DEG, differentially expressed gene; BP/CC/MF, biological process/cellular component/molecular function.

Article Snippet: Samples were fixed in 4% paraformaldehyde (PFA) at 4 °C for 30 min, permeabilized with 0.1% Triton X-100 in PBS for 15 min at room temperature, and blocked with 1% BSA in PBS for 1 h. Cells were incubated overnight at 4 °C with primary antibodies against EFNA5 (1:100; ABMART, TP72055, Shanghai, China) and EPHA4 (1:100; Proteintech, 21875-1-AP, Wuhan, China).

Techniques: In Vitro, Control

Journal: Antioxidants

Article Title: EFNA5 as an Oocyte-Derived Factor Enhances Developmental Competence by Modulating Oxidative Stress, Inflammation, and Apoptosis During In Vitro Maturation

doi: 10.3390/antiox14121476

Figure Lengend Snippet: EFNA5 improves oocyte redox homeostasis and oocyte quality. ( A ) Representative fluorescence images of intracellular GSH (left) and ROS (right) in control versus EFNA5-treated oocytes. Scale bar = 50 µm. ( B ) Quantification of relative fluorescence intensity for GSH (left) and ROS (right). ( C ) C11-BODIPY 581/591 labeling of lipid peroxidation showing reduced (red) and oxidized (green) states. Scale bar = 50 µm. ( D ) Ratio of oxidized/reduced C11-BODIPY signals. ( E ) LCA lectin staining of cortical granules in MII oocytes. Scale bar = 50 µm. ( F ) Quantification of LCA fluorescence intensity. ( G ) Immunofluorescence of γH2AX (green) indicating DNA damage (DAPI, blue). Scale bar = 20 µm. ( H ) Quantification of relative γH2AX fluorescence intensity. ( I ) qPCR analysis of antioxidant enzyme genes ( SOD1 , SOD2 , SOD3 , CATALASE , GPX ) and GSH synthesis/regeneration genes ( GCLC , GCLM , GSR ) in Control versus EFNA5-treated oocytes. Abbreviations: GSH, glutathione; ROS, reactive oxygen species; LCA, Lens culinaris agglutinin; γH2AX, phospho-H2AX (Ser139). All experiments were repeated at least three times. Data are shown as mean ± SEM. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Samples were fixed in 4% paraformaldehyde (PFA) at 4 °C for 30 min, permeabilized with 0.1% Triton X-100 in PBS for 15 min at room temperature, and blocked with 1% BSA in PBS for 1 h. Cells were incubated overnight at 4 °C with primary antibodies against EFNA5 (1:100; ABMART, TP72055, Shanghai, China) and EPHA4 (1:100; Proteintech, 21875-1-AP, Wuhan, China).

Techniques: Fluorescence, Control, Labeling, Staining, Immunofluorescence

Journal: Antioxidants

Article Title: EFNA5 as an Oocyte-Derived Factor Enhances Developmental Competence by Modulating Oxidative Stress, Inflammation, and Apoptosis During In Vitro Maturation

doi: 10.3390/antiox14121476

Figure Lengend Snippet: EFNA5 reprograms cumulus cell transcriptomes. ( A ) PCA of RNA-seq profiles from control vs. EFNA5-treated cumulus cells. ( B ) Volcano plot of DEGs; dashed lines denote thresholds |log 2 FC| = 1.5 and p -value = 0.05. Red: upregulated; blue: downregulated; gray: not significant. ( C ) Heatmap of DEGs across samples with hierarchical clustering (row-scaled Z-scores; columns: biological replicates). ( D ) Gene Ontology enrichment (BP/CC/MF). Bars indicate-log 10 (P); numbers at bar ends show gene counts. Highlighted pathways are mainly associated with inflammation and antioxidant defense. ( E ) KEGG pathway enrichment. Bubble size represents gene count; color encodes-log 10 (P); x -axis shows Gene Ratio. Highlighted pathways are mainly associated with inflammation and antioxidant defense. ( F ) Module heatmap highlighting key pathways in cumulus cells: NF-κB axis ( RELA , IKBKB , NFKBIA ), NRF2 antioxidant genes ( NRF2 , GSR , GCLC , SOD2 ), and apoptosis genes ( CASP8 , FADD , BAX , BAD ). ( G ) Adhesion-related gene heatmap (row-scaled Z-scores), including ITGAV , ITGB1 , CDH1 , CDH2 , PTK2 , FN1 , VCAN .

Article Snippet: Samples were fixed in 4% paraformaldehyde (PFA) at 4 °C for 30 min, permeabilized with 0.1% Triton X-100 in PBS for 15 min at room temperature, and blocked with 1% BSA in PBS for 1 h. Cells were incubated overnight at 4 °C with primary antibodies against EFNA5 (1:100; ABMART, TP72055, Shanghai, China) and EPHA4 (1:100; Proteintech, 21875-1-AP, Wuhan, China).

Techniques: RNA Sequencing, Control

Journal: Antioxidants

Article Title: EFNA5 as an Oocyte-Derived Factor Enhances Developmental Competence by Modulating Oxidative Stress, Inflammation, and Apoptosis During In Vitro Maturation

doi: 10.3390/antiox14121476

Figure Lengend Snippet: EFNA5 activates NRF2 signaling and suppresses NF-κB activity in COCs. ( A ) Western blot and densitometry of NRF2 in control and EFNA5 groups. Note: The blot image was horizontally flipped to align with the sample order described in the legend. ( B ) Relative mRNA levels of KEAP1 , CUL3 , and BACH1 in control and EFNA5 groups. ( C ) Western blots and densitometry of NF-κB (p65), IKBKB, and IκB. ( D ) Relative mRNA levels of IL1β , IL6 , CXCL8 , NQO1 , and TNFα in control and EFNA5 groups. All experiments were repeated at least three times. Data are shown as mean ± SEM. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Samples were fixed in 4% paraformaldehyde (PFA) at 4 °C for 30 min, permeabilized with 0.1% Triton X-100 in PBS for 15 min at room temperature, and blocked with 1% BSA in PBS for 1 h. Cells were incubated overnight at 4 °C with primary antibodies against EFNA5 (1:100; ABMART, TP72055, Shanghai, China) and EPHA4 (1:100; Proteintech, 21875-1-AP, Wuhan, China).

Techniques: Activity Assay, Western Blot, Control

Journal: Antioxidants

Article Title: EFNA5 as an Oocyte-Derived Factor Enhances Developmental Competence by Modulating Oxidative Stress, Inflammation, and Apoptosis During In Vitro Maturation

doi: 10.3390/antiox14121476

Figure Lengend Snippet: Working model of EFNA5 action in promoting oocyte developmental competence. Left, IVM without EFNA5; right, IVM with 50 ng/mL EFNA5. In cumulus cells and the oocyte, EFNA5 attenuates NF-κB signaling and its cytokine outputs ( IL-6 , TNFα , CXCL8 ), while promoting release of NRF2 from the KEAP1-CUL3-BACH1 inhibitory complex, nuclear translocation of NRF2, and transcription of antioxidant-related downstream genes ( GSH , SOD , GCLM/GCLC , CATALASE ). Solid arrows indicate activation or positive regulation, blunt-ended lines represent inhibition or suppression, and upward or downward arrows indicate increased or decreased levels, respectively. The figure was generated using Figdraw ( www.figdraw.com ).

Article Snippet: Samples were fixed in 4% paraformaldehyde (PFA) at 4 °C for 30 min, permeabilized with 0.1% Triton X-100 in PBS for 15 min at room temperature, and blocked with 1% BSA in PBS for 1 h. Cells were incubated overnight at 4 °C with primary antibodies against EFNA5 (1:100; ABMART, TP72055, Shanghai, China) and EPHA4 (1:100; Proteintech, 21875-1-AP, Wuhan, China).

Techniques: Translocation Assay, Activation Assay, Inhibition, Generated

Journal: Discover Oncology

Article Title: EFNA5 suppresses cell proliferation and tumor metastasis in hepatoma via epithelial-to-mesenchymal transition

doi: 10.1007/s12672-024-01454-7

Figure Lengend Snippet: EFNA5 expression is associated with good prognosis in hepatoma. A TCGA analysis of EFNA5 expression difference between liver tumor tissue and normal liver tissue. ** P < 0.01. B KM-plotter analysis of connection between EFNA5 expression and prognosis in hepatoma. C STRING online analysis of EFNA5 and related genes. D GO enrichment analysis of differential genes in hepatoma. E KEGG enrichment analysis of differential genes in hepatoma. F , G Representative IHC images and average density scores of EFNA5 expression in hepatoma and adjacent tissues

Article Snippet: In this experiment, the used antibodies are as follows: EFNA5 (Cat. No. ABP50058, Abbkine), GAPDH (Cat. No. AP0063, Bioworld Technology), B-actin (Cat. No. 8480, CST), EGFR (Cat. No. 51067-2-AP, PTG), E-cadherin (Cat. No. 20874-1-AP, PTG), N-cadherin (Cat. No. 13769-1-AP, PTG), c-Jun (Cat. No. 9165, CST), c-Myc (Cat. No. ab32072, Abcam), and vimentin (VIM) (Cat. No. 60330-1-Ig, PTG); HRP labeled anti-rabbit/mouse secondary antibody were purchased from Epitomics Biotechnology company.

Techniques: Expressing

Journal: Discover Oncology

Article Title: EFNA5 suppresses cell proliferation and tumor metastasis in hepatoma via epithelial-to-mesenchymal transition

doi: 10.1007/s12672-024-01454-7

Figure Lengend Snippet: EFNA5 inhibits the proliferation of hepatoma cells. A EFNA5 expression in hepatoma cell lines and normal hepatic epithelial cell line at the RNA levels by quantitative real-time PCR. *** P < 0.001. B Expression of EFNA5 in EFNA5-overexpressing HepG2 and LM3 cells, as detected by quantitative real-time PCR assays. C , D The proliferation ability of EFNA5 overexpressed hepatoma cells in LM3 and HepG2 cells was measured by CCK-8 method compared with the control group

Article Snippet: In this experiment, the used antibodies are as follows: EFNA5 (Cat. No. ABP50058, Abbkine), GAPDH (Cat. No. AP0063, Bioworld Technology), B-actin (Cat. No. 8480, CST), EGFR (Cat. No. 51067-2-AP, PTG), E-cadherin (Cat. No. 20874-1-AP, PTG), N-cadherin (Cat. No. 13769-1-AP, PTG), c-Jun (Cat. No. 9165, CST), c-Myc (Cat. No. ab32072, Abcam), and vimentin (VIM) (Cat. No. 60330-1-Ig, PTG); HRP labeled anti-rabbit/mouse secondary antibody were purchased from Epitomics Biotechnology company.

Techniques: Expressing, Real-time Polymerase Chain Reaction, CCK-8 Assay, Control

Journal: Discover Oncology

Article Title: EFNA5 suppresses cell proliferation and tumor metastasis in hepatoma via epithelial-to-mesenchymal transition

doi: 10.1007/s12672-024-01454-7

Figure Lengend Snippet: EFNA5 can inhibit the migration and invasion of hepatoma cells. A , B Representative images and quantitative analysis of cell migration based on Transwell assays. C , D The representative images and quantitative dat of the EdU assay in HepG2 and LM3

Article Snippet: In this experiment, the used antibodies are as follows: EFNA5 (Cat. No. ABP50058, Abbkine), GAPDH (Cat. No. AP0063, Bioworld Technology), B-actin (Cat. No. 8480, CST), EGFR (Cat. No. 51067-2-AP, PTG), E-cadherin (Cat. No. 20874-1-AP, PTG), N-cadherin (Cat. No. 13769-1-AP, PTG), c-Jun (Cat. No. 9165, CST), c-Myc (Cat. No. ab32072, Abcam), and vimentin (VIM) (Cat. No. 60330-1-Ig, PTG); HRP labeled anti-rabbit/mouse secondary antibody were purchased from Epitomics Biotechnology company.

Techniques: Migration, EdU Assay

Journal: Discover Oncology

Article Title: EFNA5 suppresses cell proliferation and tumor metastasis in hepatoma via epithelial-to-mesenchymal transition

doi: 10.1007/s12672-024-01454-7

Figure Lengend Snippet: EFNA5 inhibits EMT in hepatoma cells. A – C Analysis of epithelial–mesenchymal transition markers by western blotting and quantitative analysis in EFNA5 overexpression cell lysates. Overexpression of EFNA5 suppressed the protein levels of GAPDH, c-Myc, c-Jun, E-cadherin, N-cadherin, VIM, EGFR, and EFNA5 in HepG2 and LM3 cells

Article Snippet: In this experiment, the used antibodies are as follows: EFNA5 (Cat. No. ABP50058, Abbkine), GAPDH (Cat. No. AP0063, Bioworld Technology), B-actin (Cat. No. 8480, CST), EGFR (Cat. No. 51067-2-AP, PTG), E-cadherin (Cat. No. 20874-1-AP, PTG), N-cadherin (Cat. No. 13769-1-AP, PTG), c-Jun (Cat. No. 9165, CST), c-Myc (Cat. No. ab32072, Abcam), and vimentin (VIM) (Cat. No. 60330-1-Ig, PTG); HRP labeled anti-rabbit/mouse secondary antibody were purchased from Epitomics Biotechnology company.

Techniques: Western Blot, Over Expression

Journal: Discover Oncology

Article Title: EFNA5 suppresses cell proliferation and tumor metastasis in hepatoma via epithelial-to-mesenchymal transition

doi: 10.1007/s12672-024-01454-7

Figure Lengend Snippet: EFNA5 expression is associated with good prognosis in hepatoma. A TCGA analysis of EFNA5 expression difference between liver tumor tissue and normal liver tissue. ** P < 0.01. B KM-plotter analysis of connection between EFNA5 expression and prognosis in hepatoma. C STRING online analysis of EFNA5 and related genes. D GO enrichment analysis of differential genes in hepatoma. E KEGG enrichment analysis of differential genes in hepatoma. F , G Representative IHC images and average density scores of EFNA5 expression in hepatoma and adjacent tissues

Article Snippet: In line with the manufacturer’s instruction, the IHC test kit (PV-6000) for EFNA5 protein expression analysis was bought from ZsBio (Beijing, China) and used.

Techniques: Expressing

Journal: Discover Oncology

Article Title: EFNA5 suppresses cell proliferation and tumor metastasis in hepatoma via epithelial-to-mesenchymal transition

doi: 10.1007/s12672-024-01454-7

Figure Lengend Snippet: EFNA5 inhibits the proliferation of hepatoma cells. A EFNA5 expression in hepatoma cell lines and normal hepatic epithelial cell line at the RNA levels by quantitative real-time PCR. *** P < 0.001. B Expression of EFNA5 in EFNA5-overexpressing HepG2 and LM3 cells, as detected by quantitative real-time PCR assays. C , D The proliferation ability of EFNA5 overexpressed hepatoma cells in LM3 and HepG2 cells was measured by CCK-8 method compared with the control group

Article Snippet: In line with the manufacturer’s instruction, the IHC test kit (PV-6000) for EFNA5 protein expression analysis was bought from ZsBio (Beijing, China) and used.

Techniques: Expressing, Real-time Polymerase Chain Reaction, CCK-8 Assay, Control

Journal: Discover Oncology

Article Title: EFNA5 suppresses cell proliferation and tumor metastasis in hepatoma via epithelial-to-mesenchymal transition

doi: 10.1007/s12672-024-01454-7

Figure Lengend Snippet: EFNA5 can inhibit the migration and invasion of hepatoma cells. A , B Representative images and quantitative analysis of cell migration based on Transwell assays. C , D The representative images and quantitative dat of the EdU assay in HepG2 and LM3

Article Snippet: In line with the manufacturer’s instruction, the IHC test kit (PV-6000) for EFNA5 protein expression analysis was bought from ZsBio (Beijing, China) and used.

Techniques: Migration, EdU Assay

Journal: Discover Oncology

Article Title: EFNA5 suppresses cell proliferation and tumor metastasis in hepatoma via epithelial-to-mesenchymal transition

doi: 10.1007/s12672-024-01454-7

Figure Lengend Snippet: EFNA5 inhibits EMT in hepatoma cells. A – C Analysis of epithelial–mesenchymal transition markers by western blotting and quantitative analysis in EFNA5 overexpression cell lysates. Overexpression of EFNA5 suppressed the protein levels of GAPDH, c-Myc, c-Jun, E-cadherin, N-cadherin, VIM, EGFR, and EFNA5 in HepG2 and LM3 cells

Article Snippet: In line with the manufacturer’s instruction, the IHC test kit (PV-6000) for EFNA5 protein expression analysis was bought from ZsBio (Beijing, China) and used.

Techniques: Western Blot, Over Expression

Journal: Open Medicine

Article Title: Overexpression of LINC00607 inhibits cell growth and aggressiveness by regulating the miR-1289/EFNA5 axis in non-small-cell lung cancer

doi: 10.1515/med-2023-0649

Figure Lengend Snippet: EFNA5 is a downstream target of miR-1289. (a) RT-qPCR analysis of the levels of candidate mRNAs of miR-1289 in NSCLC cells after miR-1289 knockdown. (b) Western blot analysis of EFNA5 protein level in NSCLC cells after knocking down miR-1289. (c) RT-qPCR analysis of EFNA5 mRNA expression in NSCLC cells after LINC00607 overexpression. (d) Western blot analysis of EFNA5 protein level in NSCLC cells after overexpressing LINC00607. (e) Targetscan database predicted the binding site between EFNA5 and miR-1289. (f) The luciferase activity of EFNA5-WT and EFNA5-MUT affected by miR-1289 inhibition and LINC00607 silencing in NSCLC cells was assessed by the luciferase reporter assay. (g) The enrichment of LINC00607, miR-1289, and EFNA5 in anti-Ago2 and control anti-IgG groups was examined by RIP assay. (h–j) RT-qPCR and western blot analyses of EFNA5 in NSCLC cells after LINC00607 silencing and miR-1289 knockdown. ** P < 0.01, *** P < 0.001.

Article Snippet: The primary antibodies against EFNA5 (sc-81945; 1:1,000; Santa Cruz Biotechnology) and GAPDH (ab8245; 1:2,000; Abcam) were cultured with non-fat milk-blocked PVDF membrane overnight at 4°C.

Techniques: Quantitative RT-PCR, Knockdown, Western Blot, Expressing, Over Expression, Binding Assay, Luciferase, Activity Assay, Inhibition, Reporter Assay, Control

Journal: Open Medicine

Article Title: Overexpression of LINC00607 inhibits cell growth and aggressiveness by regulating the miR-1289/EFNA5 axis in non-small-cell lung cancer

doi: 10.1515/med-2023-0649

Figure Lengend Snippet: EFNA5 expression NSCLC and its association with poor prognosis. (a) RT-qPCR analysis of EFNA5 level in 35 paired NSCLC tissues and normal tissues. (b) RT-qPCR analysis of EFNA5 level in NSCLC cells and normal cells. (c) The correlation between the prognosis of NSCLC patients and EFNA5 level was analyzed through the Kaplan–Meier Plotter database. ** P < 0.01, *** P < 0.001.

Article Snippet: The primary antibodies against EFNA5 (sc-81945; 1:1,000; Santa Cruz Biotechnology) and GAPDH (ab8245; 1:2,000; Abcam) were cultured with non-fat milk-blocked PVDF membrane overnight at 4°C.

Techniques: Expressing, Quantitative RT-PCR

Journal: Open Medicine

Article Title: Overexpression of LINC00607 inhibits cell growth and aggressiveness by regulating the miR-1289/EFNA5 axis in non-small-cell lung cancer

doi: 10.1515/med-2023-0649

Figure Lengend Snippet: The influence of EFNA5 overexpression on NSCLC cell phenotypes. (a) RT-qPCR analysis of EFNA5 mRNA expression in NSCLC cells after EFNA5 overexpression. (b) Western blot analysis of EFNA5 protein level in NSCLC cells after overexpressing EFNA5. (c) MTT assay of NSCLC cell viability under EFNA5 overexpression. (d and e) Colony formation assay of NSCLC cell proliferation affected by overexpressing EFNA5. (f and g) Wound healing assay of NSCLC cell migration after overexpressing EFNA5. (h) Transwell assay of NSCLC cell invasion influenced by upregulated EFNA5. * P < 0.05, *** P < 0.001.

Article Snippet: The primary antibodies against EFNA5 (sc-81945; 1:1,000; Santa Cruz Biotechnology) and GAPDH (ab8245; 1:2,000; Abcam) were cultured with non-fat milk-blocked PVDF membrane overnight at 4°C.

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Western Blot, MTT Assay, Colony Assay, Wound Healing Assay, Migration, Transwell Assay

Journal: Open Medicine

Article Title: Overexpression of LINC00607 inhibits cell growth and aggressiveness by regulating the miR-1289/EFNA5 axis in non-small-cell lung cancer

doi: 10.1515/med-2023-0649

Figure Lengend Snippet: EFNA5 knockdown antagonizes the influence of LINC00607 on NSCLC cell phenotypes. (a) RT-qPCR and western blot analyses of EFNA5 level in NSCLC cells after downregulating EFNA5. (b) MTT assay of NSCLC cell viability in each group. (c) Colony formation assay of cell proliferative ability in each group. (d) Wound healing assay of NSCLC cell migration in each group. (e) Transwell assay of NSCLC cell invasive capability in each group. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The primary antibodies against EFNA5 (sc-81945; 1:1,000; Santa Cruz Biotechnology) and GAPDH (ab8245; 1:2,000; Abcam) were cultured with non-fat milk-blocked PVDF membrane overnight at 4°C.

Techniques: Knockdown, Quantitative RT-PCR, Western Blot, MTT Assay, Colony Assay, Wound Healing Assay, Migration, Transwell Assay

Journal: BMC Cancer

Article Title: Expression characteristic, immune signature, and prognosis value of EFNA family identified by multi-omics integrative analysis in pan-cancer

doi: 10.1186/s12885-022-09951-0

Figure Lengend Snippet: The clinical value of EFNA family in cancers. A EFNAs mRNA expression correlated with OS, DSS, DFI, and PFI from TCGA. B EFNAs mRNA expression correlated with OS and RFS in various cancers from GEO and other datasets. The prominently colored labels indicated p < 0.05. C Diagnostic ROC for EFNAs in LUAD and normal samples by TCGA combined with GTEx. D The overall survival curve of EFNA5 was analyzed integrated by the three LUAD data sets in GEO. E The nomogram for predicting 1 -, 3-and 5-year overall survival probability in LUAD patients. F Calibration of the nomogram for cox regression model and actual situation fitting analysis. G Univariate survival analysis of EFNA5 expression in subgroups with different characteristics

Article Snippet: The membrane was incubated at 4°C overnight with EFNA5 antibody (1:1000, Abmart, China) and β-tubulin antibody (Abways technology, China).

Techniques: Expressing, Diagnostic Assay

Journal: BMC Cancer

Article Title: Expression characteristic, immune signature, and prognosis value of EFNA family identified by multi-omics integrative analysis in pan-cancer

doi: 10.1186/s12885-022-09951-0

Figure Lengend Snippet: The relation between EFNA1/2/3/4/5 expression and clinical characteristics in LUAD in TCGA

Article Snippet: The membrane was incubated at 4°C overnight with EFNA5 antibody (1:1000, Abmart, China) and β-tubulin antibody (Abways technology, China).

Techniques: Expressing, Significance Assay

Journal: BMC Cancer

Article Title: Expression characteristic, immune signature, and prognosis value of EFNA family identified by multi-omics integrative analysis in pan-cancer

doi: 10.1186/s12885-022-09951-0

Figure Lengend Snippet: Univariate and multivariate analyses of overall survival in patients with LUAD in TCGA

Article Snippet: The membrane was incubated at 4°C overnight with EFNA5 antibody (1:1000, Abmart, China) and β-tubulin antibody (Abways technology, China).

Techniques:

Journal: BMC Cancer

Article Title: Expression characteristic, immune signature, and prognosis value of EFNA family identified by multi-omics integrative analysis in pan-cancer

doi: 10.1186/s12885-022-09951-0

Figure Lengend Snippet: Preliminary experimental verification of characteristics of EFNA5. A EFNA5 mRNA expression level in NSCLC cell lines from CCLE. B The expression level of EFNA5 in a normal lung bronchial epithelial cell line (HBE) and NSCLC cell lines was determined by Western Blot. C Immunocytochemistry for determining the subcellular location of EFNA5 in A431 and U251MG cell lines by HPA. EFNA5 localized to the membrane, cytosol, and vesicles (green). Microtubules are stained in red and the nucleus in blue (DAPI). D The mRNA relative expression level of EFNA5 on 13 LUAD and para-cancer paired tissues was determined by RT-qPCR. F Representative images of TMA stained with H&E and IHC for EFNA5. G EFNA5 expression levels in LUAD and peritumoral tissues determined in TMA. H EFNA5 staining score increased significantly with the characteristics of the primary tumor. I Overall survival analysis of 92 LUAD patients according to EFNA5 expression. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The membrane was incubated at 4°C overnight with EFNA5 antibody (1:1000, Abmart, China) and β-tubulin antibody (Abways technology, China).

Techniques: Expressing, Western Blot, Immunocytochemistry, Membrane, Staining, Quantitative RT-PCR

Journal: BMC Cancer

Article Title: Expression characteristic, immune signature, and prognosis value of EFNA family identified by multi-omics integrative analysis in pan-cancer

doi: 10.1186/s12885-022-09951-0

Figure Lengend Snippet: The clinical value of EFNA family in cancers. A EFNAs mRNA expression correlated with OS, DSS, DFI, and PFI from TCGA. B EFNAs mRNA expression correlated with OS and RFS in various cancers from GEO and other datasets. The prominently colored labels indicated p < 0.05. C Diagnostic ROC for EFNAs in LUAD and normal samples by TCGA combined with GTEx. D The overall survival curve of EFNA5 was analyzed integrated by the three LUAD data sets in GEO. E The nomogram for predicting 1 -, 3-and 5-year overall survival probability in LUAD patients. F Calibration of the nomogram for cox regression model and actual situation fitting analysis. G Univariate survival analysis of EFNA5 expression in subgroups with different characteristics

Article Snippet: The microarray was incubated overnight at 4°C with the indicated primary EFNA5 antibody (1:100, Abmart, China).

Techniques: Expressing, Diagnostic Assay

Journal: BMC Cancer

Article Title: Expression characteristic, immune signature, and prognosis value of EFNA family identified by multi-omics integrative analysis in pan-cancer

doi: 10.1186/s12885-022-09951-0

Figure Lengend Snippet: The relation between EFNA1/2/3/4/5 expression and clinical characteristics in LUAD in TCGA

Article Snippet: The microarray was incubated overnight at 4°C with the indicated primary EFNA5 antibody (1:100, Abmart, China).

Techniques: Expressing, Significance Assay

Journal: BMC Cancer

Article Title: Expression characteristic, immune signature, and prognosis value of EFNA family identified by multi-omics integrative analysis in pan-cancer

doi: 10.1186/s12885-022-09951-0

Figure Lengend Snippet: Univariate and multivariate analyses of overall survival in patients with LUAD in TCGA

Article Snippet: The microarray was incubated overnight at 4°C with the indicated primary EFNA5 antibody (1:100, Abmart, China).

Techniques:

Journal: BMC Cancer

Article Title: Expression characteristic, immune signature, and prognosis value of EFNA family identified by multi-omics integrative analysis in pan-cancer

doi: 10.1186/s12885-022-09951-0

Figure Lengend Snippet: Preliminary experimental verification of characteristics of EFNA5. A EFNA5 mRNA expression level in NSCLC cell lines from CCLE. B The expression level of EFNA5 in a normal lung bronchial epithelial cell line (HBE) and NSCLC cell lines was determined by Western Blot. C Immunocytochemistry for determining the subcellular location of EFNA5 in A431 and U251MG cell lines by HPA. EFNA5 localized to the membrane, cytosol, and vesicles (green). Microtubules are stained in red and the nucleus in blue (DAPI). D The mRNA relative expression level of EFNA5 on 13 LUAD and para-cancer paired tissues was determined by RT-qPCR. F Representative images of TMA stained with H&E and IHC for EFNA5. G EFNA5 expression levels in LUAD and peritumoral tissues determined in TMA. H EFNA5 staining score increased significantly with the characteristics of the primary tumor. I Overall survival analysis of 92 LUAD patients according to EFNA5 expression. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The microarray was incubated overnight at 4°C with the indicated primary EFNA5 antibody (1:100, Abmart, China).

Techniques: Expressing, Western Blot, Immunocytochemistry, Membrane, Staining, Quantitative RT-PCR

Journal: JCI Insight

Article Title: Notch-mediated Ephrin signaling disrupts islet architecture and β cell function

doi: 10.1172/jci.insight.157694

Figure Lengend Snippet: ( A ) Volcano plot showing differentially expressed genes (DEGs, adjusted P < 0.05) in islets from β-tetO-NICD and Cre + controls after 8 weeks’ Dox. ( B ) Downregulated and ( C ) upregulated biological processes in β-tetO-NICD islets, after Gene Ontology enrichment analysis using Enrichr. ( D ) Heatmap of representative DEGs across samples, with log 2 -normalized gene expression value shown. Efna5, Ephrin A5. ( E ) STRING network analysis of upregulated DEGs in β-tetO-NICD islets showing enrichment analysis of processes associated with the network, highlighting processes associated with tissue morphogenesis. ( F ) Absolute quantitation, relative to invariant control Ppia , of genes encoding Ephrin ligands in islets isolated from β-tetO-NICD and control mice after 8 weeks’ Dox ( n = 5–6 mice/group). ( G ) Representative images of pancreatic sections stained with antibodies against Efna5 and insulin from β-tetO-NICD and Cre – controls after 8 weeks’ Dox. Scale bars: 20 μm. All data are shown with group means; *, P < 0.05, **, P < 0.01, ***, P < 0.001, ****, P < 0.0001 by 2-tailed t test.

Article Snippet: We used antibodies against insulin (IR002) and somatostatin (A0566) from Dako, Agilent; Acta2 (ab5694), Adgre1 (ab6640), GFP (ab6673), and Mafa (ab26405) from Abcam; Hes1 (sc-25392) and endomucin (sc-65495) from Santa Cruz Biotechnology; Cdh1 (#3195), glucagon (#2760), and vimentin (#5741) from Cell Signaling Technology; Epcam (g8.8) from Developmental Studies Hybridoma Bank; TH (AB152) from MilliporeSigma; desmin (RB-9014-P) from Thermo Fisher Scientific; and Efna5 (AF3743), recombinant human EFNA5-Fc chimera (374-EA), and recombinant mouse TGF-β1 (7666-MB) from R&D Systems, Bio-Techne.

Techniques: Gene Expression, Quantitation Assay, Control, Isolation, Staining

Journal: JCI Insight

Article Title: Notch-mediated Ephrin signaling disrupts islet architecture and β cell function

doi: 10.1172/jci.insight.157694

Figure Lengend Snippet: ( A ) Representative images and morphometric analysis of Th-positive area in β-tetO-NICD and Cre – controls after 8 weeks’ Dox. Individual islets from at least 5 mice/group are plotted. ( B ) Experimental workflow for pseudoislet formation and cell repulsion assays. Islets were dissociated into single cells that were incubated in ultra-low-attachment wells for pseudoislet formation, or applied to coated coverslips for adhesion assays, in the presence of EFNA5-Fc chimera or Fc control. ( C ) Representative images and quantitation of pseudoislet formation of dispersed islet cells after 4-day exposure to EFNA5-Fc or Fc control, at high (55 nM) or low (13.75 mM) dose. Data pooled from 3 independent experiments. ( D ) Representative images and quantitation of β cells in EFNA5-Fc–coated condition (normalized to Fc control). Data shown as average of 3 independent experiments. Scale bars: 20 μm. All data are shown with group means; ***, P < 0.001, ****, P < 0.0001 by 2-tailed t test.

Article Snippet: We used antibodies against insulin (IR002) and somatostatin (A0566) from Dako, Agilent; Acta2 (ab5694), Adgre1 (ab6640), GFP (ab6673), and Mafa (ab26405) from Abcam; Hes1 (sc-25392) and endomucin (sc-65495) from Santa Cruz Biotechnology; Cdh1 (#3195), glucagon (#2760), and vimentin (#5741) from Cell Signaling Technology; Epcam (g8.8) from Developmental Studies Hybridoma Bank; TH (AB152) from MilliporeSigma; desmin (RB-9014-P) from Thermo Fisher Scientific; and Efna5 (AF3743), recombinant human EFNA5-Fc chimera (374-EA), and recombinant mouse TGF-β1 (7666-MB) from R&D Systems, Bio-Techne.

Techniques: Incubation, Control, Quantitation Assay

Journal: JCI Insight

Article Title: Notch-mediated Ephrin signaling disrupts islet architecture and β cell function

doi: 10.1172/jci.insight.157694

Figure Lengend Snippet: ( A ) Representative images and quantitation of pseudoislet formation of dispersed cells from human nondiabetic donor islets, after 4-day exposure to EFNA5-Fc or Fc control, at high (55 nM) or low (13.75 mM) dose. Data pooled from 3 independent experiments. ( B ) Representative images and quantitation of human β cells in EFNA5-Fc–coated condition (normalized to Fc control). Data shown as average of 3 independent experiments. Scale bars: 20 μm. All data are shown with group means; ***, P < 0.001 by 2-tailed t test.

Article Snippet: We used antibodies against insulin (IR002) and somatostatin (A0566) from Dako, Agilent; Acta2 (ab5694), Adgre1 (ab6640), GFP (ab6673), and Mafa (ab26405) from Abcam; Hes1 (sc-25392) and endomucin (sc-65495) from Santa Cruz Biotechnology; Cdh1 (#3195), glucagon (#2760), and vimentin (#5741) from Cell Signaling Technology; Epcam (g8.8) from Developmental Studies Hybridoma Bank; TH (AB152) from MilliporeSigma; desmin (RB-9014-P) from Thermo Fisher Scientific; and Efna5 (AF3743), recombinant human EFNA5-Fc chimera (374-EA), and recombinant mouse TGF-β1 (7666-MB) from R&D Systems, Bio-Techne.

Techniques: Quantitation Assay, Control

Journal: JCI Insight

Article Title: Notch-mediated Ephrin signaling disrupts islet architecture and β cell function

doi: 10.1172/jci.insight.157694

Figure Lengend Snippet: ( A ) scRNA-Seq of islets from nondiabetic and T2D patients, with β cells subclustered by metabolic pathways to define healthy (Hβ) and metabolically inflexible (MIβ) as per a previous report . tSNE, t-distributed stochastic neighbor embedding. ( B ) Violin plots of RBPJ activity in Hβ and MIβ cell populations. ( C ) GSEA of EPH-Ephrin signaling in DEGs of MIβ versus Hβ populations. NES, normalized enrichment score. ( D ) Average activity of EPH-Ephrin signaling components in Hβ and MIβ populations. ( E ) RBPJ activity as compared with EPH-Ephrin signaling components, with each dot representing a single Hβ (black) and MIβ (red) cell. Extended figure with all EPH-Ephrin signaling components and Pearson’s r values in . ( F ) Representative images of pancreatic sections stained with antibodies against EFNA5 and insulin from nondiabetic and T2D patients. Extended figure with additional staining in . Scale bars: 20 μm.

Article Snippet: We used antibodies against insulin (IR002) and somatostatin (A0566) from Dako, Agilent; Acta2 (ab5694), Adgre1 (ab6640), GFP (ab6673), and Mafa (ab26405) from Abcam; Hes1 (sc-25392) and endomucin (sc-65495) from Santa Cruz Biotechnology; Cdh1 (#3195), glucagon (#2760), and vimentin (#5741) from Cell Signaling Technology; Epcam (g8.8) from Developmental Studies Hybridoma Bank; TH (AB152) from MilliporeSigma; desmin (RB-9014-P) from Thermo Fisher Scientific; and Efna5 (AF3743), recombinant human EFNA5-Fc chimera (374-EA), and recombinant mouse TGF-β1 (7666-MB) from R&D Systems, Bio-Techne.

Techniques: Metabolic Labelling, Activity Assay, Staining