Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Sections were incubated with primary antibodies against Ki-67 (Servicebio, Cat# GB111499 ), E-cadherin (Proteintech, Cat# 20874-1-AP), and Vimentin (Proteintech, Cat# 10366-1-AP).

Techniques: Knockdown, Functional Assay, Quantitative RT-PCR, Flow Cytometry, Marker, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Derivative Assay, Co-Culture Assay, Western Blot, Migration

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 depletion in macrophages restrains LUAD growth and metastasis in vivo. (A) Schematic of orthotopic lung implantation and experimental metastasis models using LLC cells mixed with BMDMs expressing shNC or sh-circSMAD4. (B) Representative images of orthotopic lung tumors. (C) Tumor weight of orthotopic implants. (D) Overall survival of mice bearing orthotopic tumors. (E) Immunofluorescence showing F4/80 and circSMAD4 signals in tumor tissues. Scale bar, 50 μm. (F, G) Representative Ki-67 IHC staining and quantification in orthotopic tumors. Scale bar, 50 μm. (H) Representative bioluminescence images of lung tumor burden in the metastasis model. (I) Tumor weight in the metastasis model. (J) Overall survival of mice in the metastasis model. (K–M) Representative IHC staining and quantification of E-cadherin and vimentin in tumors. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Sections were incubated with primary antibodies against Ki-67 (Servicebio, Cat# GB111499 ), E-cadherin (Proteintech, Cat# 20874-1-AP), and Vimentin (Proteintech, Cat# 10366-1-AP).

Techniques: In Vivo, Expressing, Immunofluorescence, Immunohistochemistry

Journal: International Journal of Molecular Medicine

Article Title: 4-Acetylantrocamol LT3 suppresses colorectal cancer growth and metastasis via PI3K/AKT and MAPK pathway modulation

doi: 10.3892/ijmm.2026.5797

Figure Lengend Snippet: LT4 suppresses colony formation, cell migration and EMT in HCT116 cells. (A) Colony formation assay was performed after treating HCT116 cells with various concentrations of LT4 (0.1, 1, 3 and 10 μ M) for 14 days. Colonies were stained with crystal violet and quantification of colony growth was calculated relative to the vehicle-treated Ctrl. (B) Wound healing assay showing inhibition of cell migration by LT4 (0.1, 1, 3 and 10 μ M) at 24, 48 and 72 h post-scratch. Representative images and quantified healing rates are shown. Western blot analysis of EMT-related proteins (C) N-cadherin (140 kDa), (D) E-cadherin (130 kDa) and (E) Vimentin (54 kDa) in HCT116 cells after 24-h LT4 treatment (0.1, 1, 3 and 10 μ M). β-actin (45 kDa) served as the loading control. The corresponding bar graphs show semi-quantification of N-cadherin/β-actin, E-cadherin/β-actin and vimentin/β-actin expressed as fold change relative to the vehicle-treated Ctrl. Data are presented as the mean ± SD from three independent experiments (n=3). * P<0.05, ** P<0.01, *** P<0.001 vs. vehicle-treated Ctrl. LT4, 4-acetylantrocamol LT3; EMT, epithelial-mesenchymal transition; Ctrl, control.

Article Snippet: Most primary antibodies, including those against phosphorylated (p-)AKT (Ser473) (cat. no. 4060), AKT (cat. no. 9272), p-mTOR (Ser2448) (cat. no. 5536), mTOR (cat. no. 2983), p-PI3K p85 (Tyr458) (cat. no. 4228), PI3K p85 (cat. no. 4257), p-ERK1/2 (Thr202/Tyr204) (cat. no. 4370), ERK1/2 (cat. no. 4695), p-p38 (Thr180/Tyr182) (cat. no. 4511), p38 (cat. no. 9212), GSK3β (cat. no. 12456), p-GSK3β (Ser9) (cat. no. 5558), FOXO3a (cat. no. 2497), FOXO1 (cat. no. 2880), p-FOXO1 (Ser256) (cat. no. 9461), p27 kip1 (cat. no. 3686), p21 Cip1/Waf1 (cat. no. 2947), cyclooxygenase-2 (COX-2; cat. no. 12282), Bcl-2 (cat. no. 4223), Bcl-XL (cat. no. 2764), Bax (cat. no. 2772), cytochrome c oxidase subunit IV (COX IV; cat. no. 4850), β-actin (cat. no. 4970), N-cadherin (cat. no. 13116) and E-cadherin (cat. no. 3195) were purchased from Cell Signaling Technology, Inc. [rabbit host; diluted 1:1,000 in 5% BSA (Sigma-Aldrich; Merck KGaA)/PBST (PBS containing 0.1% Tween-20) with 1% NaN 3 ].

Techniques: Migration, Colony Assay, Staining, Wound Healing Assay, Inhibition, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: 4-Acetylantrocamol LT3 suppresses colorectal cancer growth and metastasis via PI3K/AKT and MAPK pathway modulation

doi: 10.3892/ijmm.2026.5797

Figure Lengend Snippet: Proposed schematic model of LT4-mediated anticancer signaling regulation in HCT116 colorectal cancer cells. This schematic summarizes the proposed major pathways and regulatory nodes modulated by LT4, based on transcriptomic, western blotting and molecular docking analyses. LT4 inhibits the PI3K/AKT/mTOR signaling cascade, as reflected by reduced phosphorylation signaling and the annotated functional outcomes (cell viability/cell proliferation and protein synthesis). In addition, LT4 shifts EMT marker expression toward an epithelial phenotype (increased E-cadherin and decreased N-cadherin). In parallel, LT4 modulates the GSK3β-FOXO1/FOXO3a node and differentially regulates MAPK signaling by suppressing ERK phosphorylation while enhancing p38 activation, accompanied by p21 upregulation, consistent with cell-cycle arrest and stress-response phenotypes. LT4 also modulates apoptosis by decreasing the levels of anti-apoptotic proteins (Bcl-2 and Bcl-XL) and increasing Bax expression, alongside mitochondrial destabilization (COX IV) and COX-2 suppression. Key hub genes identified through protein-protein interaction network analysis, SLC3A2, CCND1, PSAT1 and CHAC1, are highlighted as potential mediators linking transcriptomic regulation to these functional outcomes. Black arrows indicate canonical interactions supported by previous studies; red arrows indicate regulatory effects supported by the experimental data in the present study; dashed lines indicate docking-predicted interactions. Arrows indicate activation, whereas blunt-ended lines indicate inhibition. LT4, 4-acetylantrocamol LT3; PI3K, phosphoinositide 3-kinase; AKT, protein kinase B; mTOR, mechanistic target of rapamycin; MAPK, mitogen-activated protein kinase; ERK, extracellular signal-regulated kinase; GSK3β, glycogen synthase kinase 3 β; FOXO, forkhead box O; BAX, COX IV, cytochrome c oxidase subunit IV; COX-2, cyclooxygenase-2; N-cad, N-cadherin; E-cad, E-cadherin; EMT, epithelial-mesenchymal transition; P, phosphorylation; SLC3A2, solute carrier family 3 member 2; CCND1, cyclin D1; PSAT1, phosphoserine aminotransferase 1; CHAC1, ChaC glutathione-specific γ-glutamylcyclotransferase 1.

Article Snippet: Most primary antibodies, including those against phosphorylated (p-)AKT (Ser473) (cat. no. 4060), AKT (cat. no. 9272), p-mTOR (Ser2448) (cat. no. 5536), mTOR (cat. no. 2983), p-PI3K p85 (Tyr458) (cat. no. 4228), PI3K p85 (cat. no. 4257), p-ERK1/2 (Thr202/Tyr204) (cat. no. 4370), ERK1/2 (cat. no. 4695), p-p38 (Thr180/Tyr182) (cat. no. 4511), p38 (cat. no. 9212), GSK3β (cat. no. 12456), p-GSK3β (Ser9) (cat. no. 5558), FOXO3a (cat. no. 2497), FOXO1 (cat. no. 2880), p-FOXO1 (Ser256) (cat. no. 9461), p27 kip1 (cat. no. 3686), p21 Cip1/Waf1 (cat. no. 2947), cyclooxygenase-2 (COX-2; cat. no. 12282), Bcl-2 (cat. no. 4223), Bcl-XL (cat. no. 2764), Bax (cat. no. 2772), cytochrome c oxidase subunit IV (COX IV; cat. no. 4850), β-actin (cat. no. 4970), N-cadherin (cat. no. 13116) and E-cadherin (cat. no. 3195) were purchased from Cell Signaling Technology, Inc. [rabbit host; diluted 1:1,000 in 5% BSA (Sigma-Aldrich; Merck KGaA)/PBST (PBS containing 0.1% Tween-20) with 1% NaN 3 ].

Techniques: Western Blot, Phospho-proteomics, Functional Assay, Marker, Expressing, Activation Assay, Inhibition

Journal: Molecular & Cellular Proteomics : MCP

Article Title: O -Mannose Glycosylations Influence E-Cadherin Functional Interactions

doi: 10.1016/j.mcpro.2026.101559

Figure Lengend Snippet: Mapping the O -Man dependent E-cadherin interactome using IP screening. A , Schematic diagrams and structural model of CDH1 EC domains : ( left ) CDH1 is a transmembrane protein with five EC domains that form cis- and trans interactions; ( middle ) TMTC2 mediates O -Man on CDH1 EC B-strands, while TMTC3 mediates glycosylations on G-strands ( O -Man structures were grafted onto an AlphaFold model of EC4 using the GlycoShape tool – the mannoses are depicted as green sticks and translucent surfaces on recipient serine and threonine residues ); ( right ) schematic of the β-strand arrangement of an EC domain, highlighting O -Man sites ( green dots ) on the B- ( red ) and G- ( blue ) strands of EC2-4. B , Schematic diagram of the IP-MS-based interactome screen applied to CDH1 : Cryomilled cells are distributed to a 96-well plate and combined with different extractants; CDH1-associated complexes are affinity enriched from each extract using an antibody coupled magnetic medium and then analyzed by protein MS; the compositions of the enriched macromolecular assemblies will vary according to the stabilizing/destabilizing responses of the protein constituents and a putative interactome is constituted by the combined results. C , Results of the IP screen using 32 extraction conditions : ( upper ) silver-stained SDS-PAGE gel showing CDH1 capture by IP screening; ( lower ) hierarchical clustering of MS data, with log 2 -transformed protein abundance values from Proteome Discoverer displayed by color. Grey shading in the heatmap indicates proteins not detected (ND). Six extractants, highlighted in red, were selected for further quantitative analysis. Selected reagents present in extraction solutions are labeled with colored dots.

Article Snippet: Anti-CDH3 (MAB861, R&D systems 1:500 in 10% (v/v) FBS in PBS) and Anti-CDH1 primary antibodies (AF648 R&D systems 1:200 in 10% (v/v) FBS in PBS) were added to the cells and incubated for 1 h. The cells were washed 3 times with 100 μl of 10% (v/v) FBS in PBS and incubated with secondary antibody conjugated to fluorophore (Goat anti-Mouse IgG (H; + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 or Donkey anti-Goat IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 at 1ug/ml in 10% (v/v) FBS in PBS).

Techniques: Protein-Protein interactions, Extraction, Staining, SDS Page, Transformation Assay, Quantitative Proteomics, Labeling

Journal: Molecular & Cellular Proteomics : MCP

Article Title: O -Mannose Glycosylations Influence E-Cadherin Functional Interactions

doi: 10.1016/j.mcpro.2026.101559

Figure Lengend Snippet: I-DIRT screen for CDH1. A , Depiction of the approach : BG1 WT cells and BG1 CDH1::HA cells were cultured in both light and heavy isotope-labeling media for label-swapped I-DIRT experiments. The resulting cell powders were combined in 1:1 (w:w) ratios for IP-MS analyses. Specific CDH1 interactors are enriched in one isotope-labeled channel in MS, while non-specific interactors are quantified comparably in both the heavy and light channels. BG1 CDH1::HA cells cultured in heavy-isotope media were designated ‘I-DIRT,’ while BG1 CDH1::HA cells cultured in light-isotope media were designated ‘I-DIRT swap.’ B , Specific interactors identified across six I-DIRT experimental conditions : The interactors are grouped based on how many times they were identified as specific interactors in six extractants. Orange lines represent interactions identified in this study, while gray lines indicate interactions retrieved from the STRING database . C , overlap of the I-DIRT interactor list with two previously published datasets ( , ): the panel on the right lists the 27 common interactors. Proteins are colored by their identification frequency, as in ( B ).

Article Snippet: Anti-CDH3 (MAB861, R&D systems 1:500 in 10% (v/v) FBS in PBS) and Anti-CDH1 primary antibodies (AF648 R&D systems 1:200 in 10% (v/v) FBS in PBS) were added to the cells and incubated for 1 h. The cells were washed 3 times with 100 μl of 10% (v/v) FBS in PBS and incubated with secondary antibody conjugated to fluorophore (Goat anti-Mouse IgG (H; + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 or Donkey anti-Goat IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 at 1ug/ml in 10% (v/v) FBS in PBS).

Techniques: Cell Culture, Quantitative Proteomics, Protein-Protein interactions, Labeling

Journal: Molecular & Cellular Proteomics : MCP

Article Title: O -Mannose Glycosylations Influence E-Cadherin Functional Interactions

doi: 10.1016/j.mcpro.2026.101559

Figure Lengend Snippet: Bioinformatic analyses of CDH1 interactors. Gene Ontologies (GO) ( A – C ) and Reactome pathways ( D ) enriched among the specific CDH1 interactors. A , enrichment of GO Cellular Components (CC) associated with CDH1 interactors. B , enrichment of GO Biological Processes (BP) associated with CDH1 interactors. C , enrichment of GO Molecular Functions (MF) associated with CDH1 interactors. D , the enriched Reactome pathways is shown on the left . The specific interactors involved in each pathway are detailed on the right . E , Localizations of select CDH1 interactors : proteins with annotated localizations at the cell surface or extracellular matrix are listed. The X-axis of panels ( A – C ) represents the proportion of proteins enriched in each GO pathway.

Article Snippet: Anti-CDH3 (MAB861, R&D systems 1:500 in 10% (v/v) FBS in PBS) and Anti-CDH1 primary antibodies (AF648 R&D systems 1:200 in 10% (v/v) FBS in PBS) were added to the cells and incubated for 1 h. The cells were washed 3 times with 100 μl of 10% (v/v) FBS in PBS and incubated with secondary antibody conjugated to fluorophore (Goat anti-Mouse IgG (H; + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 or Donkey anti-Goat IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 at 1ug/ml in 10% (v/v) FBS in PBS).

Techniques:

Journal: Molecular & Cellular Proteomics : MCP

Article Title: O -Mannose Glycosylations Influence E-Cadherin Functional Interactions

doi: 10.1016/j.mcpro.2026.101559

Figure Lengend Snippet: O -Man-dependent CDH1 interactome. A , Average log 2 fold change values for CDH1 interactors in TMTC deficient cell lines: colors represent the average log 2 FC values across the six IP conditions, with red indicating increased interaction with CDH1 and blue indicating decreased interaction with CDH1. The circle size indicates the number of IP conditions where the interactor was significantly changed in different TMTC KO cell lines (log 2 FC ≥ 1 or ≤ −1, and p-adj. value ≤ 0.05). B , Selected interactors across different co-enrichment groups, for distinct IP conditions. Symbols indicate statistical significance, with “∗” representing log 2 FC ≥ 1 or ≤ −1 and p-adj. value ≤ 0.05. C , Interactions dependent on O-mannosylation position :: ( upper - panel ) example interactions affected by O -Man on EC domain G-strands; ( middle - panel ) example interactions affected by O -Man on EC domain B- and G-strands together; ( lower - panel ) ANXA1 exhibits increased co-enrichment when O -Man is depleted from EC domain B-strands (see conditions 16 and 20, KO: TMTC2 ). D , cell adhesion ability in cell lines expressing CDH1 with varying O- Man modification statuses. Data are presented as mean ± SEM ( n = 6). Statistical significance is denoted as follows: ∗∗∗ p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001.

Article Snippet: Anti-CDH3 (MAB861, R&D systems 1:500 in 10% (v/v) FBS in PBS) and Anti-CDH1 primary antibodies (AF648 R&D systems 1:200 in 10% (v/v) FBS in PBS) were added to the cells and incubated for 1 h. The cells were washed 3 times with 100 μl of 10% (v/v) FBS in PBS and incubated with secondary antibody conjugated to fluorophore (Goat anti-Mouse IgG (H; + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 or Donkey anti-Goat IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 at 1ug/ml in 10% (v/v) FBS in PBS).

Techniques: Expressing, Modification

Journal: Molecular & Cellular Proteomics : MCP

Article Title: O -Mannose Glycosylations Influence E-Cadherin Functional Interactions

doi: 10.1016/j.mcpro.2026.101559

Figure Lengend Snippet: Effects of TMTC knock - out on CDH1 and CDH3 abundance and localization. A , Western blot analysis of endogenous CDH3 abundance in BG1 cells with different TMTC KO statuses. B , Flow cytometry analysis of cell surface CDH1 and CDH3 : ( left ) representative histograms comparing fluorescence intensities in BG1 CDH1::HA cells ( green ), BG1 CDH1::HA/KO:TMTC1-4 cells ( pink ), and BG1 KO:CDH1 negative control cells ( grey ); signals normalized to mode; ( right ) Quantification of fold-change in median fluorescence intensity for surface CDH1 and CDH3 in BG1 CDH1::HA cells relative to BG1 CDH1::HA/KO:TMTC1-4 cells (n = 3). C , representative immunofluorescence images showing cellular localization of CDH1 ( green ) and CDH3 ( red ) in control BG1 CDH1::HA cells ( top panels ) and BG1 CDH1::HA/ KO :TMTC1-4 cells ( bottom panels ). Nuclei were counter-stained with DAPI ( blue ). Scale bar = 10 μm. D , Schematic model of the O-Man-dependent CDH1 interactome : some CDH1 interactors are modulated by O- Man, leading to their decreased or increased co-enrichment, based on changes e.g., in their affinity, localization, and/or abundance.

Article Snippet: Anti-CDH3 (MAB861, R&D systems 1:500 in 10% (v/v) FBS in PBS) and Anti-CDH1 primary antibodies (AF648 R&D systems 1:200 in 10% (v/v) FBS in PBS) were added to the cells and incubated for 1 h. The cells were washed 3 times with 100 μl of 10% (v/v) FBS in PBS and incubated with secondary antibody conjugated to fluorophore (Goat anti-Mouse IgG (H; + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 or Donkey anti-Goat IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 at 1ug/ml in 10% (v/v) FBS in PBS).

Techniques: Knock-Out, Western Blot, Flow Cytometry, Fluorescence, Negative Control, Immunofluorescence, Control, Staining

Journal: International Journal of Oncology

Article Title: RNA-binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability

doi: 10.3892/ijo.2026.5863

Figure Lengend Snippet: DAZAP1 induces EMT in GC cells. (A) Phase-contrast microscopy in MKN-45 cells. Scale bar, 50 μ m. (B) F-actin expression and localization were analyzed by immunocytochemistry. The cytoskeleton and nuclei were stained with rhodamine-phallotoxin (red) and Hoechst33258 (blue). (C) Immunofluorescence staining for E-cadherin (green) and vimentin (red) in MKN-45 cells. Scale bar, 20 μ m. (D) EMT biomarkers, including E-cadherin, N-cadherin and vimentin, were detected by western blotting in GC cells. (E) Representative metastatic tumors in the lungs from mice (n=4/group). Scale bar, 1 cm. (F) Number of metastatic tumors in the lung was counted. (G) Hematoxylin and eosin staining of lung sections of mice. (H) Immunohistochemical staining of vimentin. Scale bar, 100 μ m. **** P<0.001. DAZAP, deleted in azoospermia-associated protein 1; EMT, epithelial-mesenchymal transition; GC, gastric cancer; scr, scramble; si, small interfering; sh, short hairpin.

Article Snippet: Cells were incubated with primary antibodies against E-cadherin (1:200, Proteintech Group, Inc.) and Vimentin (1:200, Cell Signaling Technology, Inc.) at 4°C overnight.

Techniques: Microscopy, Expressing, Immunocytochemistry, Staining, Immunofluorescence, Western Blot, Immunohistochemical staining

Journal: International Journal of Oncology

Article Title: RNA-binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability

doi: 10.3892/ijo.2026.5863

Figure Lengend Snippet: DAZAP1 regulates NOTCH1 or JAG1 expression to affect migration/invasion via epithelial-mesenchymal transition in GC cells. The wound healing assay in MKN-45 cells confirmed that DAZAP1 regulated (A) NOTCH1(A) and JAG1. (B) Scale bar, 100 μ m. The Transwell migration assay confirmed that DAZAP1 regulates NOTCH1(C) and JAG1 (D) in MKN-45 and AGS cells, Scale bar, 150 μ m. Expression of epithelial marker E-cadherin and mesenchymal markers vimentin and N-cadherin was detected by western blotting with β-tubulin as the internal control in AGS cells. The western blotting in AGS cells confirmed that DAZAP1 regulates NOTCH1 (E) and JAG1 (F). *** P<0.01, **** P<0.001. DAZAP1, deleted in azoospermia-associated protein 1; GC, gastric cancer; scr, scramble; si, small interfering.

Article Snippet: Cells were incubated with primary antibodies against E-cadherin (1:200, Proteintech Group, Inc.) and Vimentin (1:200, Cell Signaling Technology, Inc.) at 4°C overnight.

Techniques: Expressing, Migration, Wound Healing Assay, Transwell Migration Assay, Marker, Western Blot, Control

Journal: International Journal of Oncology

Article Title: RNA-binding protein DAZAP1 promotes gastric cancer metastasis by enhancing NOTCH1 and JAG1 mRNA stability

doi: 10.3892/ijo.2026.5863

Figure Lengend Snippet: DAZAP1 promotes NOTCH1 and JAG1 expression to alter migration/invasion via epithelial-mesenchymal transition in gastric cancer cells. (A) Cell migration was tested using a monolayer wound healing assay. Scale bar, 100 μ m. (B) Migration and (C) invasion assay. Scale bar, 150 μ m. (D) Epithelial (E-cadherin) and mesenchymal markers (N-cadherin, vimentin) were detected by western blotting in AGS and MKN-45 cells. (E) Lung samples were collected from mice injected with AGS cells via the tail vein. Scale bar, 1 cm (F) Lung metastatic nodules were counted. (G) Hematoxylin and eosin and (H) vimentin staining were conducted on lung samples from these mice. Scale bar, 100 μ m. **** P<0.001. DAZAP1, deleted in azoospermia-associated protein 1; scr, scramble; si, small interfering; sh, short hairpin.

Article Snippet: Cells were incubated with primary antibodies against E-cadherin (1:200, Proteintech Group, Inc.) and Vimentin (1:200, Cell Signaling Technology, Inc.) at 4°C overnight.

Techniques: Expressing, Migration, Wound Healing Assay, Invasion Assay, Western Blot, Injection, Staining