anti phospho epha2  (Cell Signaling Technology Inc)

Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: EphA2 derived from EphA2 High cell exosomes can be internalized by EphA2 Low cells. ( A ) Expression levels of EphA2 in four GC cell lines, n = 3. ( B ) TEM characterization of exosome morphology (scale bar = 100 nm). ( C ) Particle size distribution of exosomes analyzed by NTA. ( D ) Western blot detection of exosomal markers (positive: Alix and CD9; negative: calnexin) in SGC7901 cells and exosomes. ( E ) Localization of exogenous exosomal EphA2 in MGC803 (scale bar = 25 μm). ( F ) Expression levels of EphA2 in MGC803 cells following treatment with exosomes derived from different cell sources, n = 3. (* p < 0.05, *** p < 0.001).

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Derivative Assay, Expressing, Western Blot

Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: Exosomal EphA2 derived from EphA2 High cells transforms EphA2 Low cells to promote GC progression in vitro. ( A ) Proliferation capacity of SGC7901 cells following treatment with shCtrl- or shEphA2-derived exosomes, analyzed by CCK8 assay, n = 3. ( B ) Proliferation capacity of SGC7901 cells following treatment with shCtrl- or shEphA2-derived exosomes, detected by colony formation assay, n = 3. ( C ) Migration and invasion abilities of SGC7901 cells following treatment with shCtrl- or shEphA2-derived exosomes, detected by transwell assay, n = 3 (scale bar = 200 μm). ( D ) Proliferation capacity of MGC803 cells following treatment with shCtrl- or oeEphA2-derived exosomes, analyzed by CCK8 assay, n = 3. ( E ) Proliferation capacity of MGC803 cells following treatment with shCtrl- or oeEphA2-derived exosomes, detected by colony formation assay, n = 3. ( F ) Migration and invasion abilities of MGC803 cells following treatment with shCtrl- or oeEphA2-derived exosomes, detected by transwell assay, n = 3 (scale bar = 200 μm). (ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Derivative Assay, In Vitro, CCK-8 Assay, Colony Assay, Migration, Transwell Assay

Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: Exosomal EphA2 derived from EphA2 High cells transforms EphA2 Low cells to promote GC progression in vivo. ( A ) Subcutaneous tumor formation in the left and right axillae of nude mice after injection with oeCtrl- or oeEphA2-derived exosomes ( n = 3 mice). ( B ) Comparison of tumor volume between the two groups. ( C ) Comparison of tumor weight between the two groups. ( D ) Schematic diagram of the experimental design for tail-vein injection of oeCtrl- or oeEphA2-derived exosomes in nude mice ( n = 3 mice). ( E ) In vivo imaging to detect metastatic signals in nude mice. ( F ) In vivo imaging to detect visceral metastatic foci in nude mice. ( G ) H&E staining to detect the formation of lung and liver metastatic foci after injection with oeCtrl- or oeEphA2-derived exosomes (scale bar = 100 μm). (* p < 0.05).

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Derivative Assay, In Vivo, Injection, Comparison, In Vivo Imaging, Staining

Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: Exosomal EphA2-mediated transformation of EphA2 Low to EphA2 High cells promotes GC progression through ligand-independent pathways. ( A ) Western blot analysis of EFNA1 silencing after siRNA transfection. ( B ) Transwell assay to detect changes in the migration ability of MGC803 cells after the addition of shCtrl or shEphA2 exosomes in the context of EFNA1 silencing, n = 3 (scale bar = 200 μm). ( C ) Western blot detection of the impact of exosome addition on the AKT signaling pathway. ( D ) Effect of exosome addition on the ERK signaling pathway, n = 3. ( E ) Expression of p-ERK after the addition of the ERK inhibitor U0126, n = 3. ( F ) CCK8 assay to assess the proliferation ability of MGC803 cells treated with U0126 in the presence of exosomes, n = 3. ( G ) Colony formation assay to evaluate the colony-forming ability of MGC803 cells treated with U0126 in the presence of exosomes, n = 3. ( H ) Transwell assay to measure the migration and invasion abilities of MGC803 cells treated with U0126 in the presence of exosomes, n = 3 (scale bar = 200 μm). ( I ) Expression levels of MMP2 and MMP9 in SGC7901 and MGC803 cells after EphA2 knockdown or overexpression, n = 3. ( J ) Expression levels of p-ERK and MMP2 in MGC803 cells after exosome treatment, n = 3. ( K ) Expression level of MMP2 in SGC7901 exosomes after EphA2 knockdown. (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Transformation Assay, Western Blot, Transfection, Transwell Assay, Migration, Expressing, CCK-8 Assay, Colony Assay, Knockdown, Over Expression

Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: Correlation between EphA2 expression in serum exosomes of patients with GC and tumor metastasis. ( A ) Overall survival of GC patients with high EphA2 expression ( n = 118). ( B ) Detection of EphA2 expression in human serum exosomes by ELISA ( n = 9). (* p < 0.05).

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: Schematic model illustrating how exosomal EphA2 derived from EphA2 High cells transforms EphA2 Low cells to promote gastric cancer progression in a ligand-independent manner. The figure was Created in BioRender. Zhan, L. (2026) https://BioRender.com/lsuww4f .

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Derivative Assay

Journal: Scientific Reports

Article Title: Allosteric SHP2 inhibitors suppress lung cancer cell migration by inhibiting non-canonical activation of EphA2 via the ERK-RSK signaling pathway

doi: 10.1038/s41598-025-20770-8

Figure Lengend Snippet: EphA2 phosphorylation at Ser-897 is suppressed by SHP2 inhibitors. ( A ) HeLa cells were pretreated with SHP099 (SHP, 10 µM), TNO155 (TNO, 1 µM), BI-D1870 (BI, 10 µM), or trametinib (Tram, 0.03 µM) for 1–2 h and were then stimulated with EGF (10 ng/mL), TNF-⍺ (20 ng/mL), or TPA (10 ng/mL) for 15–30 min. Whole-cell lysates were immunoblotted with EphA2, pS-EphA2, RSK1, pS-RSK1, p-ERK and β-actin antibodies. The relative quantification of pS-EphA2 shown in ( B ) is expressed as the mean ± SD of three independent experiments. P values were calculated by an ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001. ( C ) HeLa cells were pretreated with TNO155 (TNO, 1 µM) or BI-D1870 (BI, 10 µM) for 1 h and were then stimulated with TNF-⍺ (20 ng/mL) for 15 min. The expression of pS-EphA2 and EphA2 was analyzed by immunofluorescence. Representative images from three independent experiments are shown. ( D ) HeLa cells were transfected with siRNAs against GRB2 or the negative control and were then incubated for 48 h. Cells were stimulated with 20 ng/mL TNF-⍺ for 15 min. Whole-cell lysates were immunoblotted with GRB2, EphA2, pS-EphA2, and β-actin antibodies.

Article Snippet: Phospho-specific antibodies against total EphA2 (D4A2; #6997S), phospho-EphA2 (Ser-897; #6347), phospho-ERK (Thr-202/Tyr-204; #9101), phospho-p38 (Thr-180/Tyr-182; #4511S), phospho-JNK (Thr-183/Tyr-185; #4671), phospho-p65 (Ser-536; #3033), phospho-SHP2 (Tyr-542; #15543), and phospho-RSK1 (Ser-380; #11989) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Phospho-proteomics, Quantitative Proteomics, Expressing, Immunofluorescence, Transfection, Negative Control, Incubation

Journal: Scientific Reports

Article Title: Allosteric SHP2 inhibitors suppress lung cancer cell migration by inhibiting non-canonical activation of EphA2 via the ERK-RSK signaling pathway

doi: 10.1038/s41598-025-20770-8

Figure Lengend Snippet: Effects of SHP2 inhibitors on non-canonical EphA2 activation in LUAD cells. ( A ) Human lung adenocarcinoma cells (PC-9, HCC827, A925L, and A549) were treated with SHP099 (SHP, 10 µM), TNO155 (TNO, 1 µM), BI-D1870 (BI, 10 µM), or trametinib (Tram, 0.03 µM) for 1 h and were then stimulated with TNF-⍺ (20 ng/mL) for 15 min. Whole-cell lysates were immunoblotted with EphA2, pS-EphA2, RSK1, pS-RSK1, p-ERK and β-actin antibodies. ( B ) The relative quantification of pS-EphA2 shown in (A) is expressed as the mean ± SD of three independent experiments. P values were calculated by an ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001, n.s., not significant.

Article Snippet: Phospho-specific antibodies against total EphA2 (D4A2; #6997S), phospho-EphA2 (Ser-897; #6347), phospho-ERK (Thr-202/Tyr-204; #9101), phospho-p38 (Thr-180/Tyr-182; #4511S), phospho-JNK (Thr-183/Tyr-185; #4671), phospho-p65 (Ser-536; #3033), phospho-SHP2 (Tyr-542; #15543), and phospho-RSK1 (Ser-380; #11989) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Activation Assay, Quantitative Proteomics

Journal: Scientific Reports

Article Title: Allosteric SHP2 inhibitors suppress lung cancer cell migration by inhibiting non-canonical activation of EphA2 via the ERK-RSK signaling pathway

doi: 10.1038/s41598-025-20770-8

Figure Lengend Snippet: The SHP2 inhibitor suppresses the migration of lung cancer cells. ( A ) PC-9 and A549 cells were treated with SHP099 (SHP, 10 µM), TNO155 (TNO, 1 µM), BI-D1870 (BI, 10 µM), or trametinib (Tram, 0.03 µM) for 1 h. Whole-cell lysates were immunoblotted with EphA2, pS-EphA2, RSK1, p-RSK1, p-ERK and β-actin antibodies. ( B ) The relative quantification of pS-EphA2/EphA2 shown in (A) is expressed as the mean ± SD of three independent experiments. ( C , D ) A549 cells were transfected with siRNAs targeting EphA2 or the negative control RNA and subsequently subjected to a wound healing assay. A monolayer of cells was scratched mechanically and incubated for 48 h. The EphA2 expression was analyzed by immunoblotting. ( E , F ) The wound healing assay was conducted using PC-9 and A549 cells treated with TNO155 (1–3 µM) or BI-D1870 (10 µM) for 24 and 48 h, respectively. ( G , H ) The wound healing assay was conducted using A549-EphA2-KO cells treated with TNO155 (1–3 µM). ( D , F , H ) Quantification of the wound area was performed by the ImageJ software and expressed as the mean ± SD of three independent experiments. P values were calculated by an ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Phospho-specific antibodies against total EphA2 (D4A2; #6997S), phospho-EphA2 (Ser-897; #6347), phospho-ERK (Thr-202/Tyr-204; #9101), phospho-p38 (Thr-180/Tyr-182; #4511S), phospho-JNK (Thr-183/Tyr-185; #4671), phospho-p65 (Ser-536; #3033), phospho-SHP2 (Tyr-542; #15543), and phospho-RSK1 (Ser-380; #11989) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Migration, Quantitative Proteomics, Transfection, Negative Control, Wound Healing Assay, Incubation, Expressing, Western Blot, Software

Journal: Biomolecules and Biomedicine

Article Title: Pirfenidone reduces ovarian fibrosis and improves PCOS in letrozole-induced rat model

doi: 10.17305/bb.2025.12676

Figure Lengend Snippet: PFD upregulates PPAR-γ expression in the ovarian stroma of PCOS rats. (A) Representative immunohistochemical images showing PPAR-γ expression in ovarian sections. Arrows indicate PPAR-γ-positive areas in the stromal tissue surrounding the follicles. Scale bars ═ 200 µm. (B) Comparison of PPAR-γ H-scores between groups (mean ± SD; n ═ 7). * P < 0.05, **** P < 0.0001 vs Control; +++ P < 0.001, ++++ P < 0.0001 vs PCOS; ## P < 0.01 vs PCOS/PFD. PCOS: Polycystic ovarian syndrome; PFD: Pirfenidone; COC: Combined oral contraceptives; PPAR-γ: Peroxisome proliferator-activated receptor-gamma; SD: Standard deviation.

Article Snippet: Samples were then incubated overnight at 4 ∘ C with primary antibodies targeting TGFβ-1 (1:200; Santa Cruz Biotechnology, sc-130348), CTGF (1:100; BT Lab, BT-AP08174), PPAR-γ (1:200; ABclonal, A11183), MMP-9 (1:100; ABclonal, A0289), or MMP-2 (1:200; Santa Cruz Biotechnology, sc-13595).

Techniques: Expressing, Immunohistochemical staining, Comparison, Control, Standard Deviation