t24 cell line  (ATCC)

Journal: Pharmaceuticals

Article Title: Identification and Functional Analysis of Targets of Dehydrodiisoeugenol in Bladder Cancer Based on Chemoproteomics-Based Profiling

doi: 10.3390/ph19040651

Figure Lengend Snippet: DHE suppresses the proliferation and migration of T24 and 5637 bladder cancer cells. ( A ) The natural source (Myristica fragrans) and chemical structure of DHE. Atom numbering of the 2,3-dihydro-1-benzofuran core is shown for clarity; the stereogenic centers are located at C-2 and C-3. ( B , C ) Dose-response curves of T24 and 5637 cells treated with indicated concentrations of DHE for 48 h, as measured using the CCK-8 assay. ( D , E ) Proliferation curves of T24 and 5637 cells treated with DMSO or DHE (20, 40 μM) over 5 consecutive days. ( F , G ) Representative images and statistical quantification of colony formation assays for T24 ( F ) and 5637 ( G ) cells following DHE treatment. ( H ) Wound healing assays evaluating the migratory ability of T24 and 5637 cells treated with increasing concentrations of DHE (0, 10, 20, 40 μM). Representative micrographs at 0 h and 24 h are shown on the left; quantitative analysis of the area recovery percentage is shown on the right. Data are presented as Mean ± SD ( n = 3). **** p < 0.0001 vs. DMSO group ( t -test or one-way ANOVA).

Article Snippet: Cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), including the T24 cell line (ATCC Cat. No. HTB-4) and the 5637 cell line (ATCC Cat. No. HTB-9), and were cultured in RPMI-1640 medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% penicillin-streptomycin (100 U/mL; Gibco, Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Migration, CCK-8 Assay

Journal: Pharmaceuticals

Article Title: Identification and Functional Analysis of Targets of Dehydrodiisoeugenol in Bladder Cancer Based on Chemoproteomics-Based Profiling

doi: 10.3390/ph19040651

Figure Lengend Snippet: Synthesis and biological validation of a DHE-derived photoaffinity probe. ( A ) Schematic representation of the ABPP (Activity-Based Protein Profiling) workflow. The process includes cell lysis, probe incubation, UV-induced cross-linking, click chemistry-mediated biotinylation, and streptavidin-based enrichment followed by LC-MS/MS or SDS-PAGE analysis. ( B ) Synthetic route of the DHE-Probe. Reaction conditions: (a) diazirine–alkyne linker, (b) K 2 CO 3 , DMF, 65 °C, 16 h. The final probe includes a photo-cross-linker and an alkyne handle for target capturing. ( C , D ) Comparison of the anti-proliferative effects of DMSO, DHE, and DHE-Probe (40 μM) in T24 ( C ) and 5637 ( D ) bladder cancer cells. Cell viability was measured 24 h post-treatment. Data are presented as Mean ± SD ( n = 3). **** p < 0.0001 vs. DMSO group ( t -test or one-way ANOVA).

Article Snippet: Cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), including the T24 cell line (ATCC Cat. No. HTB-4) and the 5637 cell line (ATCC Cat. No. HTB-9), and were cultured in RPMI-1640 medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% penicillin-streptomycin (100 U/mL; Gibco, Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Biomarker Discovery, Derivative Assay, Activity Assay, Lysis, Incubation, Liquid Chromatography with Mass Spectroscopy, SDS Page, Comparison

Journal: Pharmaceuticals

Article Title: Identification and Functional Analysis of Targets of Dehydrodiisoeugenol in Bladder Cancer Based on Chemoproteomics-Based Profiling

doi: 10.3390/ph19040651

Figure Lengend Snippet: Gel-based ABPP validates specific protein labeling by the DHE-Probe. ( A ) Concentration-dependent labeling of the T24 cell proteome. Cell lysates were incubated with indicated concentrations of DHE-Probe (0, 5, 10, 20, 40, 80 μM), followed by UV cross-linking (365 nm) and click chemistry. The left panel shows the labeling profile (fluorescence/chemiluminescence); the right panel shows Coomassie Brilliant Blue (CBB) staining as a loading control. ( B ) Competition assay for binding specificity. Lysates were either treated with the DHE-Probe alone or pre-incubated with excess unlabeled DHE (Competition) prior to probe labeling. The significant reduction in band intensity in the competition lane (middle) confirms the specificity of the interaction. CBB staining (right) indicates equal loading. Molecular weight markers (kDa) are indicated on the left of each gel. Representative images from three independent experiments are shown.

Article Snippet: Cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), including the T24 cell line (ATCC Cat. No. HTB-4) and the 5637 cell line (ATCC Cat. No. HTB-9), and were cultured in RPMI-1640 medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% penicillin-streptomycin (100 U/mL; Gibco, Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Labeling, Concentration Assay, Incubation, Fluorescence, Staining, Control, Competitive Binding Assay, Binding Assay, Molecular Weight