Journal: Genes & Diseases

Article Title: MFAP2 promotes metastasis and drug resistance by regulating epithelial-to-mesenchymal transition through EGFR signaling pathway in colorectal cancer cells

doi: 10.1016/j.gendis.2025.101800

Figure Lengend Snippet: MFAP2 promotes epithelial–mesenchymal transition (EMT) through the EGFR-AKT-STAT3 signaling pathway in colorectal cancer (CRC) cells. (A, B) The top 20 enrichment signaling pathways regulated by MFAP2 knockdown. (C, D) VEGFR2 signaling pathway was enriched in the MFAP2 knockdown cells, shown by Gene Set Enrichment Analysis (GSEA) and essential genes in this enrichment. (E) MFAP2 knockdown affected the EGFR-AKT-STAT3 axis.

Article Snippet: Following blocking with 5% non-fat milk in PBS with 0.02% Tween 20 detergent (PBST) at room temperature for 2 h, the membranes were incubated with primary antibodies, including MFAP2 (Solarbio, China), GAPDH (BBI Co., Ltd., China), epidermal growth factor receptor (EGFR; Proteintech, China), protein kinase B (AKT) (Proteintech), signal transducer and activator of transcription 3 (STAT3) (Proteintech), and vascular endothelial growth factor A (VEGFA) (Proteintech), p-EGFR (Cell Signaling Technology, USA), p-STAT3 (Cell Signaling Technology), and p-AKT ser473 (Cell Signaling Technology) antibodies, at 4 °C overnight.

Techniques: Protein-Protein interactions, Knockdown

Journal: Journal of Extracellular Vesicles

Article Title: Extracellular Vesicle‐Mediated Nucleolin Transfer in Glioblastoma: A Targetable Axis Driving Blood‐Tumour Barrier Formation

doi: 10.1002/jev2.70268

Figure Lengend Snippet: Assembly of an MDM2‐NCL‐PROTAC‐POI quaternary complex. (A) Co‐IP of NCL, MDM2, VEGFR2, and EGFR in U87MG cells using isotype‐matched IgG or an anti‐NCL antibody. (B) Pull‐down assay of NCL, MDM2, VEGFR2, and EGFR in U87MG cells treated with 500 nM biotin‐labelled NC (Bio‐NC) or AS1411 (Bio‐AS1411) for 6 h by streptavidin‐coated magnetic beads. (C) Pull‐down assay of NCL and MDM2 in NCL siRNA (siNCL)‐transfected U87MG cells after treatment with 500 nM Bio‐AS1411 for 6 h. (D) Structure of AS1411‐Ced. (E) Structure of AS1411‐Gef. (F) Absorption spectra of NMM in the presence of 10 µM NC, AS1411, AS1411‐Ced, and AS1411‐Gef. The NMM concentration was 1 µM. (G) Co‐IP of NCL, MDM2, and VEGFR2 in U87MG cells treated with PBS or 500 nM AS1411‐Ced for 6 h using an anti‐VEGFR2 antibody in the presence of 10 µM MG132. (H) Co‐IP of NCL, MDM2, and VEGFR2 in scrambled control siRNA (siCtrl)‐ or siNCL‐transfected U87MG cells after treatment with 500 nM AS1411‐Ced for 6 h using an anti‐VEGFR2 antibody in the presence of 10 µM MG132. (I) Co‐IP of NCL, MDM2, and VEGFR2 in siCtrl or siMDM2‐transfected U87MG cells after treatment with 500 nM AS1411‐Ced for 6 h using an anti‐VEGFR2 antibody in the presence of 10 µM MG132. (J) Co‐IP of NCL, MDM2, and EGFR in U87MG cells treated with PBS or 500 nM AS1411‐Gef for 6 h using an anti‐EGFR antibody in the presence of 10 µM MG132. (K) Co‐IP of NCL, MDM2, and EGFR in siCtrl‐ or siNCL‐transfected U87MG cells after treatment with 500 nM AS1411‐Gef for 6 h using an anti‐EGFR antibody in the presence of 10 µM MG132. (L) Co‐IP of NCL, MDM2, and EGFR in siCtrl‐ or siMDM2‐transfected U87MG cells after treatment with 500 nM AS1411‐Gef for 6 h using an anti‐EGFR antibody in the presence of 10 µM MG132. Experiments were performed in triplicate and repeated three times.

Article Snippet: The lysate was centrifuged at 13,000 × g at 4°C, and the resulting supernatant was incubated overnight at 4°C with an anti‐EGFR primary antibody (1:50, CST, USA), an anti‐VEGFR2 primary antibody (1:50, CST, USA), or an anti‐NCL primary antibody (1:100, CST, USA).

Techniques: Co-Immunoprecipitation Assay, Pull Down Assay, Magnetic Beads, Transfection, Concentration Assay, Control