e cadherin  (Proteintech)

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: Cell Biology and Toxicology

Article Title: ARF6 promotes the nuclear translocation of β-catenin to facilitate AFP deficiency-mediated metastasis in CTNNB1 S33Y -mutant hepatocellular carcinoma

doi: 10.1007/s10565-026-10211-1

Figure Lengend Snippet: The transformation-promoting effect of ARF6 does not necessarily rely on AKT pathway. ( A ) Representative images of transwell migration and invasion assay on HepG2 AFP KO and HepG2 WT cells treated with MK2206 (1 μM, 48 h) or SC79 (20 μM, 1 h). 200 ×, scale bar, 50 μm. ( n = 3) ( B ) Western blot analysis of ARF6-GTP in HepG2 AFP KO and HepG2 WT cells treated with MK2206 (1 μM, 48 h). Relative protein expression was quantified ( n = 3). ( C ) Western blot analysis of ARF6-GTP in shARF6 and NC HepG2 cells treated with SC79 (20 μM, 1 h). Relative protein expression was quantified ( n = 3). ( D ) Western blot analysis of N-cadherin, E-cadherin and vimentin in HepG2 AFP KO and HepG2 WT cells treated with MK2206. Relative protein expression was quantified ( n = 3). ( E ) Western blot analysis of N-cadherin, E-cadherin and vimentin in shARF6 and NC HepG2 cells treated with SC79. Relative protein expression was quantified ( n = 3). Data are presented as the mean ± SD. And ns means no significance, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 between indicated groups

Article Snippet: To prepare antibody-bead complexes, specific primary antibodies—including those targeting β-catenin or E-cadherin— were immobilized onto protein A/G magnetic beads (MedChemExpress, HY-K0202) through a 4 h incubation at 4 ◦C with gentle rotation.

Techniques: Transformation Assay, Migration, Invasion Assay, Western Blot, Expressing

Journal: Cell Biology and Toxicology

Article Title: ARF6 promotes the nuclear translocation of β-catenin to facilitate AFP deficiency-mediated metastasis in CTNNB1 S33Y -mutant hepatocellular carcinoma

doi: 10.1007/s10565-026-10211-1

Figure Lengend Snippet: Activation of ARF6 promotes the nuclear translocation of β-catenin by assisting its dissociation from E-cadherin on the intracellular membrane. ( A ) Western blot analysis of β-catenin (β-CAT) in HepG2 AFP KO and HepG2 WT cells. ( B ) Western blot analysis of β-catenin in the cytoplasm and nucleus. Relative protein expression was quantified. ( C ) Immunofluorescence for the nuclear translocation of β-catenin. Scale bar, 10 μm. ( D ) Coimmunoprecipitation of E-cadherin and β-catenin in HepG2 AFP KO and HepG2 WT cells. Relative protein expression was quantified. ( E ) Immunofluorescence for the co-localization of β-catenin and E-cadherin in HepG2 AFP KO and HepG2 WT cells. ( F ) Western blot analysis of β-catenin in the cytoplasm, nucleus and membrane in HepG2 WT cells. treated with SecinH3(30 μM, 24 h). Relative protein expression was quantified. ( G ) Coimmunoprecipitation of E-cadherin and β-catenin, as well as ARF6 and E-cadherin in HepG2 AFP KO cells treated with SecinH3 (30 μM, 24 h). Relative protein expression was quantified. ( H ) Immunofluorescence for the co-localization of β-catenin and E-cadherin in AFP KO HepG2 cells treated with/without SecinH3(30 μM, 24 h). Scale bar, 10 μm. Data are presented as the mean ± SD ( n = 3). And ns means no significance, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 between indicated groups

Article Snippet: To prepare antibody-bead complexes, specific primary antibodies—including those targeting β-catenin or E-cadherin— were immobilized onto protein A/G magnetic beads (MedChemExpress, HY-K0202) through a 4 h incubation at 4 ◦C with gentle rotation.

Techniques: Activation Assay, Translocation Assay, Membrane, Western Blot, Expressing, Immunofluorescence

Journal: Cell Biology and Toxicology

Article Title: ARF6 promotes the nuclear translocation of β-catenin to facilitate AFP deficiency-mediated metastasis in CTNNB1 S33Y -mutant hepatocellular carcinoma

doi: 10.1007/s10565-026-10211-1

Figure Lengend Snippet: ( A , B ) IHC assay for N-cadherin, E-cadherin and Vimentin protein in liver ( A ) and lung ( B ) tissue of the metastatic HCC model mice. × 40, scale bars: 100 µm. “T” was referred to the metastatic location. ( C ) Immunofluorescence assay for β-catenin, E-cadherin protein in liver tissue of the metastatic HCC model mice. × 100, scale bars: 25 µm

Article Snippet: To prepare antibody-bead complexes, specific primary antibodies—including those targeting β-catenin or E-cadherin— were immobilized onto protein A/G magnetic beads (MedChemExpress, HY-K0202) through a 4 h incubation at 4 ◦C with gentle rotation.

Techniques: Mouse Assay, Immunofluorescence

Journal: Cell Biology and Toxicology

Article Title: ARF6 promotes the nuclear translocation of β-catenin to facilitate AFP deficiency-mediated metastasis in CTNNB1 S33Y -mutant hepatocellular carcinoma

doi: 10.1007/s10565-026-10211-1

Figure Lengend Snippet: Schematic representation of ARF6 regulations in AFP deficiency-mediated metastasis of HCC. AFP deficiency induced abnormal activation of ARF6 cells. ARF6 serves as a key pro-metastatic driver in HCC by promoting the dissociation of β-catenin from E-cadherin at the cell membrane, thereby increasing the β-catenin pool available for nuclear translocation and activation of the Wnt/β-catenin pathway. Critically, ARF6 executes this pro-metastatic program leading to enhanced migration, invasion, and EMT, only when an activating mutation CTNNB1 S33Y was harbored. Part of the drawing elements was from Biorender ( https://app.biorender.com/ )

Article Snippet: To prepare antibody-bead complexes, specific primary antibodies—including those targeting β-catenin or E-cadherin— were immobilized onto protein A/G magnetic beads (MedChemExpress, HY-K0202) through a 4 h incubation at 4 ◦C with gentle rotation.

Techniques: Activation Assay, Membrane, Translocation Assay, Migration, Mutagenesis