anti-p-epha2 (s897)  (Cell Signaling Technology Inc)

Journal: Cell Death and Differentiation

Article Title: The PLEKHA1-TACC2 fusion gene drives tumorigenesis via vascular mimicry formation in esophageal squamous-cell carcinoma

doi: 10.1038/s41418-025-01536-1

Figure Lengend Snippet: A The lysates of KYSE30 cells stably expressing Flag-tagged PLEKHA1, TACC2, Pe4Te20, Pe11Te17, Pe10Te23, Pe5Te17, Pe2Te23, or an empty vector were blotted for total EphA2, p-EphA2 (S897), total AKT, and p-AKT (S473). B The VM ability of KYSE30 cells stably expressing Flag-tagged PLEKHA1, TACC2, Pe4Te20, Pe11Te17, Pe10Te23, Pe5Te17, Pe2Te23, or an empty vector was measured by an in vitro tube formation assay. C The conditioned medium of KYSE30 cells stably expressing Flag-tagged PLEKHA1, TACC2, Pe4Te20, Pe11Te17, Pe10Te23, Pe5Te17, Pe2Te23, or an empty vector was collected for gelatin zymography (top), and the cell lysates were blotted for β-Actin (bottom). D – F Representative images of VM in tumor tissues obtained from athymic nude mice bearing KYSE30 (left) or KYSE150 (right) tumors ( D ), ESCC patients ( E ), and PDX grown in mice ( F ). Red arrows: the tubule-like VM channels. Blue arrows: the blood vessels lined by CD31-positive endothelial cells. Scale bars were indicated in the images.

Article Snippet: The antibodies utilized include the following: anti-EphA2 (1:1000; Cell Signaling Technology; 6997S); anti-β-Actin (1:4000; Cell Signaling Technology; 4970S); anti-Flag (1:1000; Proteintech; 20543-1-AP-100μl); anti-Myc (1:1000; Cell Signaling Technology; 2278S); anti-HA (1:500; Santa Cruz; sc-7392); anti-p-EphA2 (S897) (1:1000; Cell Signaling Technology; 6437S); anti-AKT (1:1000; Cell Signaling Technology; 9272S), anti-p-AKT (S473) (1:1000; Cell Signaling Technology; 4060S), anti-Nanog (1:500; Cell Signaling Technology; 4903P), anti-KLF4 (1:500; Cell Signaling Technology; 4038T), anti-SOX2 (1:500; Proteintech; 11064-1-AP-50ul) and anti-α-Tubulin (1:4000; Cell Signaling Technology; 2125S).

Techniques: Stable Transfection, Expressing, Plasmid Preparation, In Vitro, Tube Formation Assay, Zymography

Journal: Cell Death and Differentiation

Article Title: The PLEKHA1-TACC2 fusion gene drives tumorigenesis via vascular mimicry formation in esophageal squamous-cell carcinoma

doi: 10.1038/s41418-025-01536-1

Figure Lengend Snippet: A The expression of total EphA2, p-EphA2 (S897), total AKT, and p-AKT (473) in KYSE30 and KYSE150 cells stably expressing Flag-tagged Pe4Te20 treated with indicated concentrations dasatinib for 48 h was determined by immunoblotting analyses. B The VM ability of KYSE30 and KYSE150 cells stably expressing Flag-tagged Pe4Te20, Pe11Te17, Pe10Te23, Pe5Te17, Pe2Te23, without or with dasatinib (50 nM) treatment was performed using an in vitro tube formation assay. C The images of PDX G5824 and G5835 tumors from mice treated with dasatinib or vehicle for 22 days ( n = 7, each subgroup). D Growth curve of PDX G5835 (left) and G5824 (right) in mice treated with dasatinib or vehicle was plotted by measuring the relative tumor volume at indicated day. Data are presented as mean±s.e.m. The P values were calculated using the unpaired t-test. E Tumor weight of PDX G5835 (left) and G5824 (right) in mice treated with dasatinib or vehicle were measured at the endpoint of the experiment. Data are presented as mean±s.e.m. The P values were calculated using the unpaired t-test. F Tumor growth inhibition rates of dasatinib treatment on PDX G5835 and G5824. G The expression of total EphA2, p-EphA2 (S897), total AKT, and p-AKT (S473) in PDX G5835 and G5824 from mice with vehicle treatment was determined by immunoblotting analyses. H The expression of total EphA2 and p-EphA2 (S897), total AKT and p-AKT (S473) in PDX G5835 and G5824 from mice with dasatinib or vehicle treatment was determined by immunoblotting analyses. I The expression of total EphA2 and p-EphA2 (S897) in PDX G5835 and G5824 from mice with dasatinib or vehicle treatment was determined by IHC. Scale bars were indicated in the images. J H-score of EphA2 and p-EphA2 (S897) signal intensity in PDX G5835 and G5824 from mice with dasatinib or vehicle treatment as calculated by HALO analysis. The P values were calculated using one-way analysis of variance (ANOVA). K Representative images of CD31/PAS/Ter119 staining in tumor tissues of PDX G5835 and G5824 from mice with dasatinib or vehicle treatment. Red arrows: the tubule-like VM channels. Blue arrows: the blood vessels lined by CD31-positive endothelial cells. Scale bars were indicated in the images. L The number of VM channels in PDX G5835 and G5824 from mice with dasatinib or vehicle treatment as calculated by HALO analysis. The P values were calculated using one-way ANOVA. ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns indicates no significance.

Article Snippet: The antibodies utilized include the following: anti-EphA2 (1:1000; Cell Signaling Technology; 6997S); anti-β-Actin (1:4000; Cell Signaling Technology; 4970S); anti-Flag (1:1000; Proteintech; 20543-1-AP-100μl); anti-Myc (1:1000; Cell Signaling Technology; 2278S); anti-HA (1:500; Santa Cruz; sc-7392); anti-p-EphA2 (S897) (1:1000; Cell Signaling Technology; 6437S); anti-AKT (1:1000; Cell Signaling Technology; 9272S), anti-p-AKT (S473) (1:1000; Cell Signaling Technology; 4060S), anti-Nanog (1:500; Cell Signaling Technology; 4903P), anti-KLF4 (1:500; Cell Signaling Technology; 4038T), anti-SOX2 (1:500; Proteintech; 11064-1-AP-50ul) and anti-α-Tubulin (1:4000; Cell Signaling Technology; 2125S).

Techniques: Expressing, Stable Transfection, Western Blot, In Vitro, Tube Formation Assay, Inhibition, Staining

Journal: Cell Death and Differentiation

Article Title: The PLEKHA1-TACC2 fusion gene drives tumorigenesis via vascular mimicry formation in esophageal squamous-cell carcinoma

doi: 10.1038/s41418-025-01536-1

Figure Lengend Snippet: A The expression of total EphA2, p-EphA2 (S897), total AKT, and p-AKT (S473) in KYSE30 and KYSE150 cells stably expressing Flag-tagged Pe4Te20 treated with indicated concentration of regorafenib for 48 h was determined by immunoblotting analyses. B The VM ability of KYSE30 and KYSE150 cells stably expressing Flag-tagged Pe4Te20, Pe11Te17, Pe10Te23, Pe5Te17, Pe2Te23, without or with regorafenib (4 μM) treatment was performed using an in vitro tube formation assay. C The images of the tumors from the indicated transgenic mice following 4-NQO treatment first then 28 days of regorafenib or vehicle treatment ( n = 8, each subgroup). D The number of tumors from the indicated transgenic mice following 4-NQO treatment first then 28 days of regorafenib or vehicle treatment. One-way ANOVA was used for analysis. E The size of tumors from the indicated transgenic mice following 4-NQO treatment first then 28 days of regorafenib or vehicle treatment. The P values were calculated using contingency table analysis. F Tumor differentiation stages by histopathological analysis of the tongue and esophagus from the indicated transgenic mice following 4-NQO treatment first then 28 days of regorafenib or vehicle treatment. G Representative images of hematoxylin-eosin staining of tongue and esophagus obtained from the indicated transgenic mice following 4-NQO treatment first then 28 days of regorafenib or vehicle treatment. Scale bars were indicated in the images. H The expression of total EphA2 and p-EphA2 (S897) in tongue (top) and esophagus (bottom) obtained from the indicated transgenic mice following 4-NQO treatment first then 28 days of regorafenib or vehicle treatment. Scale bars were indicated in the images. I Representative images of CD31/PAS/Ter119 staining of tongue and esophagus obtained from the indicated transgenic mice following 4-NQO treatment first then 28 days of regorafenib or vehicle treatment. Red arrows: the tubule-like VM channels. Blue arrows: the blood vessels lined by CD31-positive endothelial cells. Scale bars were indicated in the images. * P < 0.05, ** P < 0.01, ns indicates no signifance.

Article Snippet: The antibodies utilized include the following: anti-EphA2 (1:1000; Cell Signaling Technology; 6997S); anti-β-Actin (1:4000; Cell Signaling Technology; 4970S); anti-Flag (1:1000; Proteintech; 20543-1-AP-100μl); anti-Myc (1:1000; Cell Signaling Technology; 2278S); anti-HA (1:500; Santa Cruz; sc-7392); anti-p-EphA2 (S897) (1:1000; Cell Signaling Technology; 6437S); anti-AKT (1:1000; Cell Signaling Technology; 9272S), anti-p-AKT (S473) (1:1000; Cell Signaling Technology; 4060S), anti-Nanog (1:500; Cell Signaling Technology; 4903P), anti-KLF4 (1:500; Cell Signaling Technology; 4038T), anti-SOX2 (1:500; Proteintech; 11064-1-AP-50ul) and anti-α-Tubulin (1:4000; Cell Signaling Technology; 2125S).

Techniques: Expressing, Stable Transfection, Concentration Assay, Western Blot, In Vitro, Tube Formation Assay, Transgenic Assay, Staining

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