rabbit monoclonal anti epha2  (Cell Signaling Technology Inc)

Journal: Cancers

Article Title: Tumor-Specific EphA2 Receptor Tyrosine Kinase Inhibits Anti-Tumor Immunity by Recruiting Suppressive Myeloid Populations in Murine Models of Non-Small Cell Lung Cancer

doi: 10.3390/cancers17162693

Figure Lengend Snippet: EphA2 confers growth advantage to NSCLC in vivo but not in vitro. ( A ) Confirmation of EphA2 overexpression in KPL and LLC cells by western blot. ( B , C ) In vitro cell viability of KPL and LLC cells with control and EphA2 overexpression by MTT and colony formation assays ( n = 4). ( D ) Representative image of bioluminescence signal in control and EphA2-overexpressing KPL tumors 14 days after subcutaneous implantation. ( E ) Representative image of bioluminescence signal 14 days after tail vein injection of control and EphA2-overexpressing KPL cells and quantification of bioluminescence signal at indicated time points (** p < 0.01, two-way ANOVA) ( F ) Representative gross specimens of GFP+ vector and EphA2-overexpressing KPL tumor-bearing lungs. ( G ) Survival of mice injected with vector or EphA2-overexpressiong KPL cells via tail vein (** p < 0.01, log-rank test). Data shown are averages ± SD.

Article Snippet: Membranes were incubated with primary monoclonal antibodies against EphA2 (Cell Signaling Technologies #6997, 1/1000, Danvers, MA, USA) and tubulin (Sigma-Aldrich #T4026, 1/2000) overnight at 4 °C, followed by three washes with TBS-T and incubation with secondary antibodies goat anti-rabbit IRDye 800CW (LI-COR #926-32211, Lincoln, NE, USA) and anti-mouse IRDye 680LT (LI-COR #926-68020, 1/20,000) for one hour at room temperature.

Techniques: In Vivo, In Vitro, Over Expression, Western Blot, Control, Injection, Plasmid Preparation

Journal: Cancers

Article Title: Tumor-Specific EphA2 Receptor Tyrosine Kinase Inhibits Anti-Tumor Immunity by Recruiting Suppressive Myeloid Populations in Murine Models of Non-Small Cell Lung Cancer

doi: 10.3390/cancers17162693

Figure Lengend Snippet: EphA2 overexpression in NSCLC does not significantly impact tumor burden or immune infiltration in nude mice. ( A ) Tumor volumes over time and weights on day 14 post-implantation of control and EphA2-overexpressing KPL subcutaneous tumors from nude mice. ( B ) Flow cytometric analysis of GFP+ KPL tumor cells and total tumor-infiltrating immune cells, as well as ( C ) tumor-infiltrating NK cells, B cells, DCs, macrophages, and Gr1+ myeloid cells. ( D ) Similar flow cytometry analysis of immune populations from draining inguinal lymph nodes. Data shown are averages ± SD ( n = 4 mice per group).

Article Snippet: Membranes were incubated with primary monoclonal antibodies against EphA2 (Cell Signaling Technologies #6997, 1/1000, Danvers, MA, USA) and tubulin (Sigma-Aldrich #T4026, 1/2000) overnight at 4 °C, followed by three washes with TBS-T and incubation with secondary antibodies goat anti-rabbit IRDye 800CW (LI-COR #926-32211, Lincoln, NE, USA) and anti-mouse IRDye 680LT (LI-COR #926-68020, 1/20,000) for one hour at room temperature.

Techniques: Over Expression, Control, Flow Cytometry

Journal: Cancers

Article Title: Tumor-Specific EphA2 Receptor Tyrosine Kinase Inhibits Anti-Tumor Immunity by Recruiting Suppressive Myeloid Populations in Murine Models of Non-Small Cell Lung Cancer

doi: 10.3390/cancers17162693

Figure Lengend Snippet: EphA2 overexpression in NSCLC decreases lymphocytic and increases myeloid infiltrate in tumor-bearing lungs. ( A ) Representative flow cytometry plots and quantification of GFP+ KPL and immune cells from vector control and EphA2-overexpressing tumor-bearing lungs on day 14 post-tail vein injection. ( B ) Similar flow plots and analysis of CD4+ and CD8+ T cells and NK cells, as well as ( C ) quantification of DCs, macrophages, and monocytes. Data shown are averages ± SD ( n = 3 mice per group, * p < 0.05; ** p < 0.01; *** p < 0.001, two-tailed unpaired Student’s t test with Welch correction).

Article Snippet: Membranes were incubated with primary monoclonal antibodies against EphA2 (Cell Signaling Technologies #6997, 1/1000, Danvers, MA, USA) and tubulin (Sigma-Aldrich #T4026, 1/2000) overnight at 4 °C, followed by three washes with TBS-T and incubation with secondary antibodies goat anti-rabbit IRDye 800CW (LI-COR #926-32211, Lincoln, NE, USA) and anti-mouse IRDye 680LT (LI-COR #926-68020, 1/20,000) for one hour at room temperature.

Techniques: Over Expression, Flow Cytometry, Plasmid Preparation, Control, Injection, Two Tailed Test

Journal: Cancers

Article Title: Tumor-Specific EphA2 Receptor Tyrosine Kinase Inhibits Anti-Tumor Immunity by Recruiting Suppressive Myeloid Populations in Murine Models of Non-Small Cell Lung Cancer

doi: 10.3390/cancers17162693

Figure Lengend Snippet: EphA2 overexpression in NSCLC suppresses tumor-infiltrating T cells. ( A ) Lung weights and quantification of GFP+ KPL cells via flow cytometry from vector control and EphA2-overexpressing tumor-bearing lungs with equalized tumor burden. ( B ) Flow cytometric analysis of total immune cells, CD4+ and CD8+ T cells, and NK cells in KPL tumor-bearing lungs. ( C ) Representative flow histograms of CD44 and CD69 expression on CD8 T cells and quantification of CD44, CD69, and CD25 activation markers on CD4 and CD8 T cells. ( D ) Quantification of PD-1 and CTLA-4 exhaustion markers on CD4+ and CD8+ T cells. Data shown are averages ± SD ( n = 3–6 mice per group, * p < 0.05; ** p < 0.01, two-tailed unpaired Student’s t test with Welch correction).

Article Snippet: Membranes were incubated with primary monoclonal antibodies against EphA2 (Cell Signaling Technologies #6997, 1/1000, Danvers, MA, USA) and tubulin (Sigma-Aldrich #T4026, 1/2000) overnight at 4 °C, followed by three washes with TBS-T and incubation with secondary antibodies goat anti-rabbit IRDye 800CW (LI-COR #926-32211, Lincoln, NE, USA) and anti-mouse IRDye 680LT (LI-COR #926-68020, 1/20,000) for one hour at room temperature.

Techniques: Over Expression, Flow Cytometry, Plasmid Preparation, Control, Expressing, Activation Assay, Two Tailed Test

Journal: Cancers

Article Title: Tumor-Specific EphA2 Receptor Tyrosine Kinase Inhibits Anti-Tumor Immunity by Recruiting Suppressive Myeloid Populations in Murine Models of Non-Small Cell Lung Cancer

doi: 10.3390/cancers17162693

Figure Lengend Snippet: Gene expression profiling reveals higher expression of myeloid markers and chemoattractants in EphA2-overexpressing tumors. ( A ) Average pathway scores of vector control and EphA2-overexpressing KPL tumors calculated from normalized gene expression data using nanoString nSolver software. ( B ) Comparison of cancer progression and macrophage functions pathway scores between control and EphA2-overexpressing samples. ( n = 6 mice per group, ** p < 0.01, unpaired Mann-Whitney test) ( C ) Volcano plot of statistically significant differentially expressed genes. ( D ) Heatmap depicting standardized expression of differentially expressed myeloid markers (green bar), myeloid-attracting chemokines (pink bar), and immunosuppressive proteins (yellow bar). ( E ) RT-PCR validation of nanoString hits. ( n = 6 mice per group, * p < 0.05, one-sample Wilcoxon signed rank test). Data shown are averages ± SD.

Article Snippet: Membranes were incubated with primary monoclonal antibodies against EphA2 (Cell Signaling Technologies #6997, 1/1000, Danvers, MA, USA) and tubulin (Sigma-Aldrich #T4026, 1/2000) overnight at 4 °C, followed by three washes with TBS-T and incubation with secondary antibodies goat anti-rabbit IRDye 800CW (LI-COR #926-32211, Lincoln, NE, USA) and anti-mouse IRDye 680LT (LI-COR #926-68020, 1/20,000) for one hour at room temperature.

Techniques: Gene Expression, Expressing, Plasmid Preparation, Control, Software, Comparison, MANN-WHITNEY, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery

Journal: Virulence

Article Title: Mannan is a context-dependent shield that modifies virulence in Nakaseomyces glabratus .

doi: 10.1080/21505594.2025.2491650

Figure Lengend Snippet: Figure 3. N. glabratus mnn10Δ mutants induce higher phosphorylation of OEC EphA2 compared to their respective wild-type parental strains. Western blotting for total and phosphorylated EphA2 (pEpha2). Human α-actin, loading control. Figure is representative of three independent experiments. OECs, oral epithelial cells, negative control. SC5314, C. albicans reference strain, positive control. WT, wild-type strain. mnn10Δ, null mutant strain.

Article Snippet: Antibodies for assessment of EphA2 phosphorylation Anti-EphA2 (D4A2) and anti-p-EphA2 (S897) rabbit monoclonal antibodies were purchased from Cell Signaling Technologies (CST Inc., MA, USA) (catalogue numbers 6997S and 6347S, respectively).

Techniques: Phospho-proteomics, Western Blot, Control, Negative Control, Positive Control, Mutagenesis