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rabbit monoclonal anti py588 epha2 (R&D Systems)

Name: rabbit monoclonal anti py588 epha2
Brand: R&D Systems
Rating: 93 (15 reviews)

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R&D Systems rabbit monoclonal anti py588 epha2

A) Analysis of log 2 transformed FPKM gene expression values extracted from bulk RNA seq published previously (Smith R. et al. Scientific Reports 2020 ) shows high <t>EphA2</t> expression in castration resistant, commercially available human PCa cell lines. B) Immunoblot characterization of select human PCa cell lines show increased EphA2 expression and S897 phosphorylation in mCRPC derived cell lines. C) EphA2 expression in representative human prostate cancer cell lines. D) Lysate of human prostate cancer tissues reveal increased expression of EphA2 in bone metastasis samples. Lysates were prepared from micro-dissected human prostate cancer tissues under the Rapid Autopsy Program at the Univ. Mich. PCa SPORE.

Rabbit Monoclonal Anti Py588 Epha2, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit monoclonal anti py588 epha2/product/R&D Systems
Average 93 stars, based on 15 article reviews

rabbit monoclonal anti py588 epha2 - by Bioz Stars, 2026-02

93/100 stars

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1) Product Images from "Skeletal Metastasis of Prostate Cancer Is Augmented by Activation of EphA2 Noncanonical Signaling and Ligand-Deficient Bone Microenvironment"

Article Title: Skeletal Metastasis of Prostate Cancer Is Augmented by Activation of EphA2 Noncanonical Signaling and Ligand-Deficient Bone Microenvironment

Journal: bioRxiv

doi: 10.1101/2024.09.25.615079

Figure Legend Snippet: A) Analysis of log 2 transformed FPKM gene expression values extracted from bulk RNA seq published previously (Smith R. et al. Scientific Reports 2020 ) shows high EphA2 expression in castration resistant, commercially available human PCa cell lines. B) Immunoblot characterization of select human PCa cell lines show increased EphA2 expression and S897 phosphorylation in mCRPC derived cell lines. C) EphA2 expression in representative human prostate cancer cell lines. D) Lysate of human prostate cancer tissues reveal increased expression of EphA2 in bone metastasis samples. Lysates were prepared from micro-dissected human prostate cancer tissues under the Rapid Autopsy Program at the Univ. Mich. PCa SPORE.

Techniques Used: Transformation Assay, Expressing, RNA Sequencing Assay, Western Blot, Derivative Assay

A) Diagram depicting the serial propagation of TRAMP-C1 cells in syngeneic mice to select for cells with increased tumorigenic capacity. B) The in vivo selection of cells resulted in accelerated tumor growth and reduced latency. C) Immunoblot analyses of the in vivo selected (IV1) and the parental TRAMP-C1 cells using the indicated antibodies. D,E,F,G,H,I,J) Quantification of relative band intensity (n = 4). All bands normalized to tubulin, except for pY588 and pS897 which are normalized to total EphA2 expression. K) Immunoblot of TRAMP-C1 parental and IV1 cells after stimulation with Ephrin-A1-Fc ligand for the indicated times. L,M) MPC3 and MycCaP cells were propagated in syngeneic C57Bl/6 and FVB mice, respectively. Lysates of the parental and the in vivo selected cells (IV1) were subject to immunoblot using the indicated antibodies.

Figure Legend Snippet: A) Diagram depicting the serial propagation of TRAMP-C1 cells in syngeneic mice to select for cells with increased tumorigenic capacity. B) The in vivo selection of cells resulted in accelerated tumor growth and reduced latency. C) Immunoblot analyses of the in vivo selected (IV1) and the parental TRAMP-C1 cells using the indicated antibodies. D,E,F,G,H,I,J) Quantification of relative band intensity (n = 4). All bands normalized to tubulin, except for pY588 and pS897 which are normalized to total EphA2 expression. K) Immunoblot of TRAMP-C1 parental and IV1 cells after stimulation with Ephrin-A1-Fc ligand for the indicated times. L,M) MPC3 and MycCaP cells were propagated in syngeneic C57Bl/6 and FVB mice, respectively. Lysates of the parental and the in vivo selected cells (IV1) were subject to immunoblot using the indicated antibodies.

Techniques Used: In Vivo, Selection, Western Blot, Expressing

A) Immunohistochemical (IHC) analyses of tissue microarray (TMA) prepared from primary, lymph node metastatic (LN), and bone metastatic human PCa tissues from the Rapid Autopsy Program of the Pacific Northwest PCa SPORE spotted. B and C) The relative IHC staining strengths of EphA2 and pS897-EphA2 on TMA was scored using 0-3 scale (0, absent; 1, weak; 2, moderate; 3, strong) . Data consists of 27 bone metastatic samples, and 18 soft tissue samples including 10 lymph node, 5 liver, and 3 lung metastatic samples. Comparisons were made using an unpaired t-test (B, p = 0.0003 C, p < 0.0001) D) mRNA expression analysis of human PCa metastatic biopsies revealed that bone metastases have the highest EphA2 expression compared to other sites of disease dissemination. Data from 208 samples from the SU2C/PCF Dream Team. mRNA Expression z-scores relative to all samples (log FPKM Capture) accessed via cBioportal. E) Kaplan-Meier curve of patients with bone metastases according to the levels of EphA2 expression.

Figure Legend Snippet: A) Immunohistochemical (IHC) analyses of tissue microarray (TMA) prepared from primary, lymph node metastatic (LN), and bone metastatic human PCa tissues from the Rapid Autopsy Program of the Pacific Northwest PCa SPORE spotted. B and C) The relative IHC staining strengths of EphA2 and pS897-EphA2 on TMA was scored using 0-3 scale (0, absent; 1, weak; 2, moderate; 3, strong) . Data consists of 27 bone metastatic samples, and 18 soft tissue samples including 10 lymph node, 5 liver, and 3 lung metastatic samples. Comparisons were made using an unpaired t-test (B, p = 0.0003 C, p < 0.0001) D) mRNA expression analysis of human PCa metastatic biopsies revealed that bone metastases have the highest EphA2 expression compared to other sites of disease dissemination. Data from 208 samples from the SU2C/PCF Dream Team. mRNA Expression z-scores relative to all samples (log FPKM Capture) accessed via cBioportal. E) Kaplan-Meier curve of patients with bone metastases according to the levels of EphA2 expression.

Techniques Used: Immunohistochemical staining, Microarray, Immunohistochemistry, Expressing

A) Immunoblot analysis of PC3 cells transduced with lentivirus to express WT-EphA2 or S897A-EphA2. Empty vector-transduced cells were used as control. Cells were starved for 24 hours in serum-free medium and stimulated with either FBS alone or FBS togethr with ephrin-A1-Fc. B) PC3 cells overexpressing WT-EphA2 or S897A-EphA2 mutant were xenografted subcutaneously into nude mice (n = 10 per group). Vector alone cells were used as control. Comparison between WT-EphA2 and S897A-EphA2 made with an unpaired t-test at day 46 post-injection (p = 0.0402).

Figure Legend Snippet: A) Immunoblot analysis of PC3 cells transduced with lentivirus to express WT-EphA2 or S897A-EphA2. Empty vector-transduced cells were used as control. Cells were starved for 24 hours in serum-free medium and stimulated with either FBS alone or FBS togethr with ephrin-A1-Fc. B) PC3 cells overexpressing WT-EphA2 or S897A-EphA2 mutant were xenografted subcutaneously into nude mice (n = 10 per group). Vector alone cells were used as control. Comparison between WT-EphA2 and S897A-EphA2 made with an unpaired t-test at day 46 post-injection (p = 0.0402).

Techniques Used: Western Blot, Transduction, Plasmid Preparation, Control, Mutagenesis, Comparison, Injection

Figure Legend Snippet: A) Analysis of log 2 transformed FPKM gene expression values extracted from bulk RNA seq published previously (Smith R. et al. Scientific Reports 2020 ) showing low Efna1-5 gene expression in commercially available castration resistant and androgen sensitive human PCa cell lines. B) Immunoblot characterization of select PCa cell lines reveals simultaneous high EphA2 expression and low EphrinA1 expression in metastatic castration resistant cells compared with androgen sensitive cells. C ) Quantitative analysis of data in (B). D) Kaplan-Meier survival curve from the MSK dataset ( Cancer Cell 2010 ). Samples were aggregated into the top 25 percent EFNA1 expressing ( EFNA1 High), and lower 75 percent EFNA1 expressing ( EFNA1 Low). Data were accessed via cBioportal.

Techniques Used: Transformation Assay, Expressing, RNA Sequencing Assay, Western Blot

A) Lysates of PC-3 cells transduced by lentiviral vector expressing EFNA1 (EA1) or empty control vector were subject to immunoblot with the indicated antibodies. B,C) Quantification of pY588 marking canonical signaling and pS897 marking noncanonical signaling by EphA2, relative to total EphA2 expression. D) Vec. and EA1-expressing PC-3 cells were stimulated with recombinant exogenous EA1-Fc ligand in vitro for the indicated times, and lysates were subject to immunoblot with the indicated antibodies. E) PY99, a monoclonal antibody that recognizes most pY motifs, was used to probe overall pY profiles using lysates from Vec. and EA1-expressing PC-3 cells stimulated with EA1-Fc. N-cadherin and pY416-Src were also probed. F) PC-3 cells expressing EA1 exhibit reduced cell growth in an in vitro cell proliferation assay. G) Vec. control and EA1-expressing cells were implanted subcutaneously into nude mice. Paired t-test to be used to monitor (p = 0.0319). G) Bioluminescence imaging of mice four and six weeks after intratibial injection of PC3-vec. or PC-3-EA1 expressing cells tagged with luciferase. H) X-ray imaging of mice 6 weeks post intratibial injection of PC3-Vec and PC3-EA1 cells.

Figure Legend Snippet: A) Lysates of PC-3 cells transduced by lentiviral vector expressing EFNA1 (EA1) or empty control vector were subject to immunoblot with the indicated antibodies. B,C) Quantification of pY588 marking canonical signaling and pS897 marking noncanonical signaling by EphA2, relative to total EphA2 expression. D) Vec. and EA1-expressing PC-3 cells were stimulated with recombinant exogenous EA1-Fc ligand in vitro for the indicated times, and lysates were subject to immunoblot with the indicated antibodies. E) PY99, a monoclonal antibody that recognizes most pY motifs, was used to probe overall pY profiles using lysates from Vec. and EA1-expressing PC-3 cells stimulated with EA1-Fc. N-cadherin and pY416-Src were also probed. F) PC-3 cells expressing EA1 exhibit reduced cell growth in an in vitro cell proliferation assay. G) Vec. control and EA1-expressing cells were implanted subcutaneously into nude mice. Paired t-test to be used to monitor (p = 0.0319). G) Bioluminescence imaging of mice four and six weeks after intratibial injection of PC3-vec. or PC-3-EA1 expressing cells tagged with luciferase. H) X-ray imaging of mice 6 weeks post intratibial injection of PC3-Vec and PC3-EA1 cells.

Techniques Used: Plasmid Preparation, Expressing, Control, Western Blot, Recombinant, In Vitro, Proliferation Assay, Imaging, Injection, Luciferase

Image Search Results

Journal: bioRxiv

Article Title: Skeletal Metastasis of Prostate Cancer Is Augmented by Activation of EphA2 Noncanonical Signaling and Ligand-Deficient Bone Microenvironment

doi: 10.1101/2024.09.25.615079

Figure Lengend Snippet: A) Analysis of log 2 transformed FPKM gene expression values extracted from bulk RNA seq published previously (Smith R. et al. Scientific Reports 2020 ) shows high EphA2 expression in castration resistant, commercially available human PCa cell lines. B) Immunoblot characterization of select human PCa cell lines show increased EphA2 expression and S897 phosphorylation in mCRPC derived cell lines. C) EphA2 expression in representative human prostate cancer cell lines. D) Lysate of human prostate cancer tissues reveal increased expression of EphA2 in bone metastasis samples. Lysates were prepared from micro-dissected human prostate cancer tissues under the Rapid Autopsy Program at the Univ. Mich. PCa SPORE.

Article Snippet: Other antibodies were purchased as follows: Rabbit monoclonal anti-GAPDH (Cell Signaling Technology #2118S), rabbit monoclonal anti-pS473-Akt (Cell Signaling Technology #4060), rabbit polyclonal anti-pErk1/pErk2 (Cell Signaling Technology #9101S), rabbit monoclonal anti-EphA2 (Cell Signaling Technology #6697S), rat monoclonal anti-EphA2-PE (R&D Systems FAB639P), rabbit monoclonal anti-AR (Abcam ab133273), rabbit polyclonal anti-pY772-EphA2 (Cell Signaling Technology #8244S), rabbit monoclonal anti-pY588-EphA2 (Cell Signaling Technology #12677S), mouse monoclonal anti-Tubulin (R&D Systems MAB9344) Goat anti-rabbit and goat anti-mouse secondary antibodies conjugated to horseradish peroxidase (HRP) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA) and from Biorad (#1706515, and #1706516 respectively).

Techniques: Transformation Assay, Expressing, RNA Sequencing Assay, Western Blot, Derivative Assay

Journal: bioRxiv

Article Title: Skeletal Metastasis of Prostate Cancer Is Augmented by Activation of EphA2 Noncanonical Signaling and Ligand-Deficient Bone Microenvironment

doi: 10.1101/2024.09.25.615079

Figure Lengend Snippet: A) Diagram depicting the serial propagation of TRAMP-C1 cells in syngeneic mice to select for cells with increased tumorigenic capacity. B) The in vivo selection of cells resulted in accelerated tumor growth and reduced latency. C) Immunoblot analyses of the in vivo selected (IV1) and the parental TRAMP-C1 cells using the indicated antibodies. D,E,F,G,H,I,J) Quantification of relative band intensity (n = 4). All bands normalized to tubulin, except for pY588 and pS897 which are normalized to total EphA2 expression. K) Immunoblot of TRAMP-C1 parental and IV1 cells after stimulation with Ephrin-A1-Fc ligand for the indicated times. L,M) MPC3 and MycCaP cells were propagated in syngeneic C57Bl/6 and FVB mice, respectively. Lysates of the parental and the in vivo selected cells (IV1) were subject to immunoblot using the indicated antibodies.

Techniques: In Vivo, Selection, Western Blot, Expressing

Journal: bioRxiv

Article Title: Skeletal Metastasis of Prostate Cancer Is Augmented by Activation of EphA2 Noncanonical Signaling and Ligand-Deficient Bone Microenvironment

doi: 10.1101/2024.09.25.615079

Figure Lengend Snippet: A) Immunohistochemical (IHC) analyses of tissue microarray (TMA) prepared from primary, lymph node metastatic (LN), and bone metastatic human PCa tissues from the Rapid Autopsy Program of the Pacific Northwest PCa SPORE spotted. B and C) The relative IHC staining strengths of EphA2 and pS897-EphA2 on TMA was scored using 0-3 scale (0, absent; 1, weak; 2, moderate; 3, strong) . Data consists of 27 bone metastatic samples, and 18 soft tissue samples including 10 lymph node, 5 liver, and 3 lung metastatic samples. Comparisons were made using an unpaired t-test (B, p = 0.0003 C, p < 0.0001) D) mRNA expression analysis of human PCa metastatic biopsies revealed that bone metastases have the highest EphA2 expression compared to other sites of disease dissemination. Data from 208 samples from the SU2C/PCF Dream Team. mRNA Expression z-scores relative to all samples (log FPKM Capture) accessed via cBioportal. E) Kaplan-Meier curve of patients with bone metastases according to the levels of EphA2 expression.

Techniques: Immunohistochemical staining, Microarray, Immunohistochemistry, Expressing

Journal: bioRxiv

Article Title: Skeletal Metastasis of Prostate Cancer Is Augmented by Activation of EphA2 Noncanonical Signaling and Ligand-Deficient Bone Microenvironment

doi: 10.1101/2024.09.25.615079

Figure Lengend Snippet: A) Immunoblot analysis of PC3 cells transduced with lentivirus to express WT-EphA2 or S897A-EphA2. Empty vector-transduced cells were used as control. Cells were starved for 24 hours in serum-free medium and stimulated with either FBS alone or FBS togethr with ephrin-A1-Fc. B) PC3 cells overexpressing WT-EphA2 or S897A-EphA2 mutant were xenografted subcutaneously into nude mice (n = 10 per group). Vector alone cells were used as control. Comparison between WT-EphA2 and S897A-EphA2 made with an unpaired t-test at day 46 post-injection (p = 0.0402).

Techniques: Western Blot, Transduction, Plasmid Preparation, Control, Mutagenesis, Comparison, Injection

Journal: bioRxiv

Article Title: Skeletal Metastasis of Prostate Cancer Is Augmented by Activation of EphA2 Noncanonical Signaling and Ligand-Deficient Bone Microenvironment

doi: 10.1101/2024.09.25.615079

Figure Lengend Snippet: A) Analysis of log 2 transformed FPKM gene expression values extracted from bulk RNA seq published previously (Smith R. et al. Scientific Reports 2020 ) showing low Efna1-5 gene expression in commercially available castration resistant and androgen sensitive human PCa cell lines. B) Immunoblot characterization of select PCa cell lines reveals simultaneous high EphA2 expression and low EphrinA1 expression in metastatic castration resistant cells compared with androgen sensitive cells. C ) Quantitative analysis of data in (B). D) Kaplan-Meier survival curve from the MSK dataset ( Cancer Cell 2010 ). Samples were aggregated into the top 25 percent EFNA1 expressing ( EFNA1 High), and lower 75 percent EFNA1 expressing ( EFNA1 Low). Data were accessed via cBioportal.

Techniques: Transformation Assay, Expressing, RNA Sequencing Assay, Western Blot

Journal: bioRxiv

Article Title: Skeletal Metastasis of Prostate Cancer Is Augmented by Activation of EphA2 Noncanonical Signaling and Ligand-Deficient Bone Microenvironment

doi: 10.1101/2024.09.25.615079

Figure Lengend Snippet: A) Lysates of PC-3 cells transduced by lentiviral vector expressing EFNA1 (EA1) or empty control vector were subject to immunoblot with the indicated antibodies. B,C) Quantification of pY588 marking canonical signaling and pS897 marking noncanonical signaling by EphA2, relative to total EphA2 expression. D) Vec. and EA1-expressing PC-3 cells were stimulated with recombinant exogenous EA1-Fc ligand in vitro for the indicated times, and lysates were subject to immunoblot with the indicated antibodies. E) PY99, a monoclonal antibody that recognizes most pY motifs, was used to probe overall pY profiles using lysates from Vec. and EA1-expressing PC-3 cells stimulated with EA1-Fc. N-cadherin and pY416-Src were also probed. F) PC-3 cells expressing EA1 exhibit reduced cell growth in an in vitro cell proliferation assay. G) Vec. control and EA1-expressing cells were implanted subcutaneously into nude mice. Paired t-test to be used to monitor (p = 0.0319). G) Bioluminescence imaging of mice four and six weeks after intratibial injection of PC3-vec. or PC-3-EA1 expressing cells tagged with luciferase. H) X-ray imaging of mice 6 weeks post intratibial injection of PC3-Vec and PC3-EA1 cells.

Techniques: Plasmid Preparation, Expressing, Control, Western Blot, Recombinant, In Vitro, Proliferation Assay, Imaging, Injection, Luciferase

EPHA2 is expressed on the cell surface of undifferentiated mouse ESCs and downregulated during differentiation. (A) Relative gene expression of Epha2 mRNA in mouse ESCs (D3) cultured without LIF for 0, 3, 5, and 7 days. (B) Immunoblotting of EPHA2 protein of mouse ESCs cultured as in (A). (C) EPHA2 protein levels in (B) normalized to α-Tubulin. (D) Immunofluorescent staining of mouse ESCs. The cells were cultured with or without LIF for 7 days. Bars; 200 μm. (E, F) Flow cytometric analysis of EPHA2 protein on the cell surface of mouse ESCs. Living cells were stained with an EPHA2 antibody. The propidium iodide + dead cells were removed from the analysis. FSC and SSC profiles (E) and the histogram of EPHA2-AF488 levels (F) of the gated cells in (E) were shown. Statistical analyses in (A) and (C) were calculated by Dunnett’s test. Data were graphed as means ± SE of 3 independent biological replicates. * P < .05; ** P < .01; *** P < .001.

Journal: Stem Cells Translational Medicine

Article Title: EPHA2 is a novel cell surface marker of OCT4-positive undifferentiated cells during the differentiation of mouse and human pluripotent stem cells

doi: 10.1093/stcltm/szae036

Figure Lengend Snippet: EPHA2 is expressed on the cell surface of undifferentiated mouse ESCs and downregulated during differentiation. (A) Relative gene expression of Epha2 mRNA in mouse ESCs (D3) cultured without LIF for 0, 3, 5, and 7 days. (B) Immunoblotting of EPHA2 protein of mouse ESCs cultured as in (A). (C) EPHA2 protein levels in (B) normalized to α-Tubulin. (D) Immunofluorescent staining of mouse ESCs. The cells were cultured with or without LIF for 7 days. Bars; 200 μm. (E, F) Flow cytometric analysis of EPHA2 protein on the cell surface of mouse ESCs. Living cells were stained with an EPHA2 antibody. The propidium iodide + dead cells were removed from the analysis. FSC and SSC profiles (E) and the histogram of EPHA2-AF488 levels (F) of the gated cells in (E) were shown. Statistical analyses in (A) and (C) were calculated by Dunnett’s test. Data were graphed as means ± SE of 3 independent biological replicates. * P < .05; ** P < .01; *** P < .001.

Article Snippet: For immunofluorescent analyses of human iPSCs, mouse monoclonal anti-human EPHA2 (MAB3035; R&D systems) and goat polyclonal anti-human SOX17 (AF1924, R&D systems) were used.

Techniques: Expressing, Cell Culture, Western Blot, Staining

Knock-down of Epha2 induces spontaneous differentiation of mouse ESCs. (A) qRT-PCR analysis of Epha2 mRNA after knock-down in mouse ESCs (D3). The cells were infected with Epha2 shRNA retrovirus and cultured with or without LIF for 7 days. (B) Images of phase contrast and alkaline phosphatase (AP) staining of Epha2 KD mouse ESCs cultured in ES maintenance medium with LIF for 5 days. Bars; 200 μm. (C) qRT-PCR analysis of undifferentiated state-specific marker genes in Epha2 KD mouse ESCs. (D) Immunofluorescent staining of Epha2 KD mouse ESCs cultured with LIF for 7 days. Bars; 100 μm. (E) Culture conditions of Epha2 KD mouse ESCs after infection with Epha2 shRNA retrovirus. One day after infection with Epha2 shRNA virus, mouse ESCs were cultured with or without 2i in the presence of G418 and LIF for 4 days. (F) Phase contrast images of Epha2 KD mouse ESCs cultured as in (E). Bar; 200 μm. (G) qRT-PCR analysis of Epha2 and undifferentiated state-specific marker genes in Epha2 KD mouse ESCs cultured with 2i. All qRT-PCR analyses were performed with 3 independent biological replicates and graphed as means ± SE. The significant differences were calculated by Tukey test in (A and C) and t -test in (G). * P < .05; ** P < .01, *** P < .001.

Journal: Stem Cells Translational Medicine

Article Title: EPHA2 is a novel cell surface marker of OCT4-positive undifferentiated cells during the differentiation of mouse and human pluripotent stem cells

doi: 10.1093/stcltm/szae036

Figure Lengend Snippet: Knock-down of Epha2 induces spontaneous differentiation of mouse ESCs. (A) qRT-PCR analysis of Epha2 mRNA after knock-down in mouse ESCs (D3). The cells were infected with Epha2 shRNA retrovirus and cultured with or without LIF for 7 days. (B) Images of phase contrast and alkaline phosphatase (AP) staining of Epha2 KD mouse ESCs cultured in ES maintenance medium with LIF for 5 days. Bars; 200 μm. (C) qRT-PCR analysis of undifferentiated state-specific marker genes in Epha2 KD mouse ESCs. (D) Immunofluorescent staining of Epha2 KD mouse ESCs cultured with LIF for 7 days. Bars; 100 μm. (E) Culture conditions of Epha2 KD mouse ESCs after infection with Epha2 shRNA retrovirus. One day after infection with Epha2 shRNA virus, mouse ESCs were cultured with or without 2i in the presence of G418 and LIF for 4 days. (F) Phase contrast images of Epha2 KD mouse ESCs cultured as in (E). Bar; 200 μm. (G) qRT-PCR analysis of Epha2 and undifferentiated state-specific marker genes in Epha2 KD mouse ESCs cultured with 2i. All qRT-PCR analyses were performed with 3 independent biological replicates and graphed as means ± SE. The significant differences were calculated by Tukey test in (A and C) and t -test in (G). * P < .05; ** P < .01, *** P < .001.

Article Snippet: For immunofluorescent analyses of human iPSCs, mouse monoclonal anti-human EPHA2 (MAB3035; R&D systems) and goat polyclonal anti-human SOX17 (AF1924, R&D systems) were used.

Techniques: Knockdown, Quantitative RT-PCR, Infection, shRNA, Cell Culture, Staining, Marker, Virus

$Expression of EPHA2 in heterogenous subpopulation of human PSCs. (A) qRT-PCR analysis of EPHA2 and undifferentiated state-specific marker genes during EB-based random differentiation of human iPSCs (201B7) without basic FGF. Statistical analysis was done by Dunnett’s test comparing to day 0 and graphed as means ± SE of 3 independent experiments. * P < .05; ** P < .01. (B) Representative flow cytometric plots of living human iPSCs stained with EPHA2 antibody-conjugated with AFF488. Gate3 and Gate4 were sorted as EPHA2 − and EPHA2 + cell populations, respectively. See also . (C) Feature plots of EPHA2 and undifferentiated state-specific genes in publicly available undifferentiated human ESC H1 and H9 data subsets from GSE75748 scRNA-seq dataset. Note that EPHA2 expression in hESCs was heterogeneous. Normalized expression levels were plotted. (D, E) Immunofluorescent staining of human iPSC cultured on SyntheMax II-coated plate with StemFit medium. The cells were fixed with paraformaldehyde in PBS and permeabilized. Bars; 200 μm. (F) qRT-PCR analysis of fractioned EPHA2 + and EPHA2 - subpopulations. EPHA2 + cells express higher OCT4 and NANOG , than EPHA2 − cells. Means ± SE of 3 independent experiments were shown. Statistical significance was defined as * P < .05 by t -test.$

Journal: Stem Cells Translational Medicine

Article Title: EPHA2 is a novel cell surface marker of OCT4-positive undifferentiated cells during the differentiation of mouse and human pluripotent stem cells

doi: 10.1093/stcltm/szae036

Figure Lengend Snippet: Expression of EPHA2 in heterogenous subpopulation of human PSCs. (A) qRT-PCR analysis of EPHA2 and undifferentiated state-specific marker genes during EB-based random differentiation of human iPSCs (201B7) without basic FGF. Statistical analysis was done by Dunnett’s test comparing to day 0 and graphed as means ± SE of 3 independent experiments. * P < .05; ** P < .01. (B) Representative flow cytometric plots of living human iPSCs stained with EPHA2 antibody-conjugated with AFF488. Gate3 and Gate4 were sorted as EPHA2 − and EPHA2 + cell populations, respectively. See also . (C) Feature plots of EPHA2 and undifferentiated state-specific genes in publicly available undifferentiated human ESC H1 and H9 data subsets from GSE75748 scRNA-seq dataset. Note that EPHA2 expression in hESCs was heterogeneous. Normalized expression levels were plotted. (D, E) Immunofluorescent staining of human iPSC cultured on SyntheMax II-coated plate with StemFit medium. The cells were fixed with paraformaldehyde in PBS and permeabilized. Bars; 200 μm. (F) qRT-PCR analysis of fractioned EPHA2 + and EPHA2 - subpopulations. EPHA2 + cells express higher OCT4 and NANOG , than EPHA2 − cells. Means ± SE of 3 independent experiments were shown. Statistical significance was defined as * P < .05 by t -test.

Article Snippet: For immunofluorescent analyses of human iPSCs, mouse monoclonal anti-human EPHA2 (MAB3035; R&D systems) and goat polyclonal anti-human SOX17 (AF1924, R&D systems) were used.

Techniques: Expressing, Quantitative RT-PCR, Marker, Staining, Cell Culture

Transplantation of EPHA2 + cells into immune-deficient mice formed tumors in vivo. (A) Immunofluorescent staining of mouse EBs differentiated into hepatocyte lineages. Expression of an early hepatocyte marker AFP at day 10 and a mature marker ALB at day 14 were detected. Bars; 200 μm. (B) Depletion of undifferentiated ES colonies after removal of EPHA2 + cells from EBs. Oct4-egfp ESCs were differentiated by EB formation for 10 and 14 days. EPHA2 + cells were removed from EBs using anti-EPHA2 antibody-bound MACS after dissociation with trypsin/EDTA. The residual cells were cultured in ES maintenance medium with LIF for 7 days. The alkaline phosphatase (AP) activity was visualized by incubating with AP substrate. Bar; 2 cm. (C) The number of EGFP + cell colonies in (B). Statistical analysis was done by Tukey test and graphed as means ± SE of 4 independent experiments. (D) Scheme of in vivo transplantation experiment. Mouse ESCs (D3) were differentiated into hepatocyte linages and the EBs were dissociated by EDTA treatment. The cells were transplanted into SCID mice after depletion of EPHA2 + cells by MACS. (E) Decreased teratoma formation after transplantation of EPHA2 − cells. White arrowheads indicate teratomas. (F) H&E staining of teratomas formed in the testicular subcutaneous tissue without MACS procedure. Typical cell types of 3 germ layers were confirmed. Bar; 500 μm. (G) Typical teratoma formation after transplantation of dissociated EB at day 10 through hepatic portal vein. White arrowheads indicate teratomas. (H) Quantification of teratoma formation in (E) and (G). Statistical analysis was done by Chi-square test, * P < .05, ** P < .01.

Journal: Stem Cells Translational Medicine

Article Title: EPHA2 is a novel cell surface marker of OCT4-positive undifferentiated cells during the differentiation of mouse and human pluripotent stem cells

doi: 10.1093/stcltm/szae036

Figure Lengend Snippet: Transplantation of EPHA2 + cells into immune-deficient mice formed tumors in vivo. (A) Immunofluorescent staining of mouse EBs differentiated into hepatocyte lineages. Expression of an early hepatocyte marker AFP at day 10 and a mature marker ALB at day 14 were detected. Bars; 200 μm. (B) Depletion of undifferentiated ES colonies after removal of EPHA2 + cells from EBs. Oct4-egfp ESCs were differentiated by EB formation for 10 and 14 days. EPHA2 + cells were removed from EBs using anti-EPHA2 antibody-bound MACS after dissociation with trypsin/EDTA. The residual cells were cultured in ES maintenance medium with LIF for 7 days. The alkaline phosphatase (AP) activity was visualized by incubating with AP substrate. Bar; 2 cm. (C) The number of EGFP + cell colonies in (B). Statistical analysis was done by Tukey test and graphed as means ± SE of 4 independent experiments. (D) Scheme of in vivo transplantation experiment. Mouse ESCs (D3) were differentiated into hepatocyte linages and the EBs were dissociated by EDTA treatment. The cells were transplanted into SCID mice after depletion of EPHA2 + cells by MACS. (E) Decreased teratoma formation after transplantation of EPHA2 − cells. White arrowheads indicate teratomas. (F) H&E staining of teratomas formed in the testicular subcutaneous tissue without MACS procedure. Typical cell types of 3 germ layers were confirmed. Bar; 500 μm. (G) Typical teratoma formation after transplantation of dissociated EB at day 10 through hepatic portal vein. White arrowheads indicate teratomas. (H) Quantification of teratoma formation in (E) and (G). Statistical analysis was done by Chi-square test, * P < .05, ** P < .01.

Article Snippet: For immunofluorescent analyses of human iPSCs, mouse monoclonal anti-human EPHA2 (MAB3035; R&D systems) and goat polyclonal anti-human SOX17 (AF1924, R&D systems) were used.

Techniques: Transplantation Assay, In Vivo, Staining, Expressing, Marker, Cell Culture, Activity Assay

Co-expression of EPHA2 with OCT4 in EBs during human iPSC differentiation into hepatocyte. (A, B) Relative gene expression of undifferentiation and differentiation markers during hepatic induction. Statistical analysis was done by Dunnett’s test against day 0 and graphed as means ± SE of 3 independent biological replicates. * P < .05; ** P < .01, *** P < .001. (C) Immunofluorescent staining of EBs at days 5, 8, and 10. Bars; 200 μm. (D) Enlarged images of EB at day 5 in (C). Bars; 50 μm. (E, F) Quantification of immune-positive cells in . Box plot showing the percentage of EPHA2 + cells among SOX17 + or OCT4 + cells (E). Box plot showing the percentage of EPHA2 + and TRA1-81 + cells among OCT4 + cells (F). Each box represents 1st quartile, median, and 3rd quartile, and whiskers show the minimum and maximum values. Ten images of independent EBs were analyzed. Total count of DAPI + nuclei per image were between 1 × 10 3 and 2 × 10 3 . Statistical significance was defined by Tukey test of SOX17 and OCT4, respectively in (E) and t -test between EPHA2 and TRA1-81 in (F). **P p < .01, *** P < .001, N.S; no significance between 3 with P > .05.

Journal: Stem Cells Translational Medicine

Article Title: EPHA2 is a novel cell surface marker of OCT4-positive undifferentiated cells during the differentiation of mouse and human pluripotent stem cells

doi: 10.1093/stcltm/szae036

Figure Lengend Snippet: Co-expression of EPHA2 with OCT4 in EBs during human iPSC differentiation into hepatocyte. (A, B) Relative gene expression of undifferentiation and differentiation markers during hepatic induction. Statistical analysis was done by Dunnett’s test against day 0 and graphed as means ± SE of 3 independent biological replicates. * P < .05; ** P < .01, *** P < .001. (C) Immunofluorescent staining of EBs at days 5, 8, and 10. Bars; 200 μm. (D) Enlarged images of EB at day 5 in (C). Bars; 50 μm. (E, F) Quantification of immune-positive cells in . Box plot showing the percentage of EPHA2 + cells among SOX17 + or OCT4 + cells (E). Box plot showing the percentage of EPHA2 + and TRA1-81 + cells among OCT4 + cells (F). Each box represents 1st quartile, median, and 3rd quartile, and whiskers show the minimum and maximum values. Ten images of independent EBs were analyzed. Total count of DAPI + nuclei per image were between 1 × 10 3 and 2 × 10 3 . Statistical significance was defined by Tukey test of SOX17 and OCT4, respectively in (E) and t -test between EPHA2 and TRA1-81 in (F). **P p < .01, *** P < .001, N.S; no significance between 3 with P > .05.

Article Snippet: For immunofluorescent analyses of human iPSCs, mouse monoclonal anti-human EPHA2 (MAB3035; R&D systems) and goat polyclonal anti-human SOX17 (AF1924, R&D systems) were used.

Techniques: Expressing, Staining

Figure 1. Immunohistochemical staining of glioma tissues. (A) MMP‑2 and (B) EphA2 expression was identified by immunofluorescence staining in the cytoplasm of glioma cells. Positive cells were stained brown. Percentages of stained cells are presented in the bar graphs. Data represent the mean ± standard deviation (n=5). ***P<0.001. Scale bar, 30 µm. EphA2, ephrin type‑A receptor 2; MMP‑2, metalloproteinase 2.

Journal: Oncology letters

Article Title: The combined use of EphA2/MMP-2 expression and MRI findings contributes to the determination of cerebral glioma grade.

doi: 10.3892/ol.2019.10912

Figure Lengend Snippet: Figure 1. Immunohistochemical staining of glioma tissues. (A) MMP‑2 and (B) EphA2 expression was identified by immunofluorescence staining in the cytoplasm of glioma cells. Positive cells were stained brown. Percentages of stained cells are presented in the bar graphs. Data represent the mean ± standard deviation (n=5). ***P<0.001. Scale bar, 30 µm. EphA2, ephrin type‑A receptor 2; MMP‑2, metalloproteinase 2.

Article Snippet: One slide was routinely stained with hematoxylin and eosin in 25 ̊C for 1‐5 min. After blocking in BSA solution 37 ̊C for 30 min (cat. no. P0260; Beyotime Institute of Biotechnology), the other slide was used for IHC staining using mouse anti-human MMP-2 and mouse anti-human EphA2 (Boster cat. no. M00286 and BM0833, both 1:200) monoclonal antibodies detected by streptavidin-peroxidase or 3'-diaminobenzidine visualization kits (OriGene Technologies, Inc.).

Techniques: Immunohistochemical staining, Staining, Expressing, Immunofluorescence, Standard Deviation

Figure 3. Pearson's correlation analysis of the magnetic resonance imaging parameters and immunohistochemistry results. Scatterplots presenting the correla- tion between (A) MMP‑2 and (B) EphA2 positivity with the EI; (C) MMP‑2 and (D) EphA2 positivity with the EP; and (E) MMP‑2 and (F) EphA2 positivity with maximum tumor diameter. EI, edema index; EP, enhancement percentage; EphA2, ephrin type‑A receptor 2; MMP‑2, metalloproteinase 2.

Journal: Oncology letters

Article Title: The combined use of EphA2/MMP-2 expression and MRI findings contributes to the determination of cerebral glioma grade.

doi: 10.3892/ol.2019.10912

Figure Lengend Snippet: Figure 3. Pearson's correlation analysis of the magnetic resonance imaging parameters and immunohistochemistry results. Scatterplots presenting the correla- tion between (A) MMP‑2 and (B) EphA2 positivity with the EI; (C) MMP‑2 and (D) EphA2 positivity with the EP; and (E) MMP‑2 and (F) EphA2 positivity with maximum tumor diameter. EI, edema index; EP, enhancement percentage; EphA2, ephrin type‑A receptor 2; MMP‑2, metalloproteinase 2.

Techniques: Magnetic Resonance Imaging, Immunohistochemistry

Journal: Journal for Immunotherapy of Cancer

Article Title: Safety, tolerability, pharmacokinetics, and pharmacodynamics of the afucosylated, humanized anti-EPHA2 antibody DS-8895a: a first-in-human phase I dose escalation and dose expansion study in patients with advanced solid tumors

doi: 10.1186/s40425-019-0679-9

Figure Lengend Snippet: Patient characteristics

Article Snippet: EPHA2 was detected using anti-human EPHA2 mouse monoclonal antibodies (clones 018 and 058, Daiichi Sankyo Co., Ltd.).

Techniques: Expressing

Journal: Journal for Immunotherapy of Cancer

doi: 10.1186/s40425-019-0679-9

Figure Lengend Snippet: Efficacy results

Article Snippet: EPHA2 was detected using anti-human EPHA2 mouse monoclonal antibodies (clones 018 and 058, Daiichi Sankyo Co., Ltd.).

Techniques:

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1) Product Images from "Skeletal Metastasis of Prostate Cancer Is Augmented by Activation of EphA2 Noncanonical Signaling and Ligand-Deficient Bone Microenvironment"

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