hek293t cells  (ATCC)

Journal: International Journal of Molecular Medicine

Article Title: Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2-positive breast cancer

doi: 10.3892/ijmm.2026.5751

Figure Lengend Snippet: EBA downregulates HER2, p95HER2, HER3 and AKT expression. (A) Immunoblot analysis of HER2, p95HER2 and p-HER2 (Y1221/1222) in JIMT-1 cells treated with EBA for 48 h. (B) Immunoblot analysis of HER3, p-HER3 (Y1289), AKT and p-AKT following treatment with EBA (48 h) in JIMT-1 cells. (C) Immunoblot analysis of HER2, HER3 and EGFR following IP with anti-HER2 antibody in JIMT-1 cells treated with EBA. In silico molecular docking of EBA with the crystal structure of HER2-KD. (D) Surface map of lipophilic and hydrophilic properties at the ATP-binding site of HER2-KD (red, hydrophobic; blue, hydrophilic). (E) 2D interaction diagram showing intermolecular interactions between EBA and HER2-KD. Key amino acid residues within the binding pocket are shown. (F) Predicted binding pose of EBA (purple stick model) within the tyrosine kinase domain of HER2 (blue ribbon). (G) 293T cells were treated with DMSO or EBA for 1 h at 37°C, followed by heating for 3 min. Soluble fractions were collected following centrifugation and analyzed by immunoblotting using an anti-HER2 antibody. EBA, ebastine; p-, phosphorylated; IP, immunoprecipitation; IB, immunoblotting; KD, kinase domain PCB, protein complex binding; TM, transmembrane; a.a., amino acid.

Article Snippet: 293T (American Type Culture Collection) cells were cultured overnight at 37°C in a humidified atmosphere with 5% CO 2 , and treated with either DMSO (vehicle) or 30 μ M ebastine for 1 h at 37°C.

Techniques: Expressing, Western Blot, In Silico, Binding Assay, Centrifugation, Immunoprecipitation

Journal: Biochemistry and Biophysics Reports

Article Title: Ectopic endometrial mesenchymal stem cell-derived exosomal miR-4466 promotes angiogenesis by targeting RUNX1 in adenomyosis

doi: 10.1016/j.bbrep.2026.102457

Figure Lengend Snippet: miR-4466 directly regulates RUNX1 expression in HUVECs. (A) A Venn diagram shows target genes of miR-4466 predicted by the miRDB, miRWalk, miRPathDB, mirDIP databases. (B) Schematic diagram illustrating the predicted miR-4466 binding sites with the 3′ UTR of RUNX1. (C, D) Relative expression of target genes was detected by qRT-PCR in HUVECs (C) and HEK-293T cells (D) transfected with NC-mimics and miR-mimics. n = 3. E, F Relative expression of target genes was detected by qRT-PCR in HUVECs (E) and HEK-293T cells (F) transfected with NC-inhibitor and miR-inhibitor. n = 3. (G) Schematic representation of the wild-type (Wt) and mutant-type (Mut) binding site between the 3′ UTR of RUNX1 and miR-4466. (H) Relative luciferase activity of reporter constructs containing miRNA binding sites transfected with NC-mimics and miR-mimics in HUVECs. n = 3. (I) Western blot of RUNX1 and VEGFA protein expression in HUVECs after transfection with NC-mimics/inhibitor or miR-mimics/inhibitor for 48 h (Full-length blot are presented in Supplementary Figure). (J, K) Protein levels of RUNX1 (J) and VEGFA (K) were normalized to GAPDH protein expression level in (I). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 and ns: P > 0.05.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) and human embryonic kidney 293T (HEK-293T) cells were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Expressing, Binding Assay, Quantitative RT-PCR, Transfection, Mutagenesis, Luciferase, Activity Assay, Construct, Western Blot