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: bioRxiv

Article Title: Preferential Invasion of Differentiated Bladder Carcinoma Cells by Flagellated Group B2 Escherichia Coli

doi: 10.64898/2026.04.16.718932

Figure Lengend Snippet: Invasion of E. coli strains from three phylogenetic groups in HTB-9 and CRL2169. Group AD refers to strains from either phylogenetic group A or D. IBC (intracellular bacterial community) refers to intracellular bacteria recovered. (A) Invasion into HTB-9: colonies recovered. (B) Invasion into HTB-9: cumulative comparison of group B2 vs non-B2 strains. This panel replots all the technical replicates for each strain from panel A. (C) Invasion into CRL2169: colonies recovered. (D) Invasion into CRL2169: cumulative comparison of group B2 vs non-B2 strains. This panel replots all the technical replicates for each strain from panel C. (E) Ratio of invasion into HTB-9 to CRL2169: individual strains. (E) Ratio of invasion into HTB-9 to CRL2169: cumulative comparison.

Article Snippet: The grade 2 male urothelial carcinoma cell line, HTB-9 (also called line 5637, ATCC-5637) and the grade 1 female urothelial carcinoma cell line, CRL-2169 (ATCC-SW 780) were grown in Lebovitz’s 15 (L-15) (ATCC-30-2008) medium supplemented with antibiotic-antimycotic (Sigma-A5955) and 10% FBS (Thermo-A3160401) at 37°C to roughly 80% confluency.

Techniques: Bacteria, Comparison

Journal: bioRxiv

Article Title: Preferential Invasion of Differentiated Bladder Carcinoma Cells by Flagellated Group B2 Escherichia Coli

doi: 10.64898/2026.04.16.718932

Figure Lengend Snippet: Comparison of wild-type UTI89 and mutants in the pili major structural subunit, fimA , the flagellar major subunit, fliC , and a double mutant in both fimA and fliC . Statistical significance was determined by Kruskal-Wallis non-parametric test of multiple comparisons. (A) Invasion into CRL2169. For parental, Δ fimA , Δ fliC , and Δ fimA Δ fliC strains, the samples sizes were 5, 3, 6, and 6, respectively. For the double mutant, three replicates had no recoverable bacteria. (B) Invasion into HTB-9. For parental UTI89 and Δ fimA , Δ fliC , and Δ fimA Δ fliC derivatives, the samples sizes were 6, 3, 6, and 6, respectively. For the double mutant, none of the six replicates had recoverable bacteria. (C) Mouse model of urinary tract infection. For parental, Δ fimA , Δ fliC , and Δ fimA Δ fliC strains, the samples sizes were 23, 8, 12, and 8, respectively. For the double mutant, none of the 8 replicates had recoverable bacteria.

Article Snippet: The grade 2 male urothelial carcinoma cell line, HTB-9 (also called line 5637, ATCC-5637) and the grade 1 female urothelial carcinoma cell line, CRL-2169 (ATCC-SW 780) were grown in Lebovitz’s 15 (L-15) (ATCC-30-2008) medium supplemented with antibiotic-antimycotic (Sigma-A5955) and 10% FBS (Thermo-A3160401) at 37°C to roughly 80% confluency.

Techniques: Comparison, Mutagenesis, Bacteria, Infection

Journal: bioRxiv

Article Title: Preferential Invasion of Differentiated Bladder Carcinoma Cells by Flagellated Group B2 Escherichia Coli

doi: 10.64898/2026.04.16.718932

Figure Lengend Snippet: Invasion of a hyperflagellated derivative of W3110 in HTB-9 and CRL2169. No bacterial strain was considered invasive. ns, not significant

Article Snippet: The grade 2 male urothelial carcinoma cell line, HTB-9 (also called line 5637, ATCC-5637) and the grade 1 female urothelial carcinoma cell line, CRL-2169 (ATCC-SW 780) were grown in Lebovitz’s 15 (L-15) (ATCC-30-2008) medium supplemented with antibiotic-antimycotic (Sigma-A5955) and 10% FBS (Thermo-A3160401) at 37°C to roughly 80% confluency.

Techniques:

Journal: bioRxiv

Article Title: Preferential Invasion of Differentiated Bladder Carcinoma Cells by Flagellated Group B2 Escherichia Coli

doi: 10.64898/2026.04.16.718932

Figure Lengend Snippet: Overview of endocytosis and IBC formation and ratio of endocytic proteins and receptors. (A) Overview of endocytosis. Red and blue receptors are TLR5 and TLR4, respectively. Both TLRs are shown binding their activating signals, flagella and LPS, denoted by the black flagella and purple triangle. Green angles represent both clathrin and caveolin. Image was generated using Biorender.com. (B) Uroplakin transcript levels between CRL2169 and HTB-9 from public databases. (C) TLR-2, -4, and -5 transcript levels from public databases. (D) Transcript ratios for endocytosis genes between CRL2169 and HTB-9 from ( , ).

Article Snippet: The grade 2 male urothelial carcinoma cell line, HTB-9 (also called line 5637, ATCC-5637) and the grade 1 female urothelial carcinoma cell line, CRL-2169 (ATCC-SW 780) were grown in Lebovitz’s 15 (L-15) (ATCC-30-2008) medium supplemented with antibiotic-antimycotic (Sigma-A5955) and 10% FBS (Thermo-A3160401) at 37°C to roughly 80% confluency.

Techniques: Binding Assay, Generated

Journal: bioRxiv

Article Title: Preferential Invasion of Differentiated Bladder Carcinoma Cells by Flagellated Group B2 Escherichia Coli

doi: 10.64898/2026.04.16.718932

Figure Lengend Snippet: Transcript ratios of TLRs from HTB-9 to CRL2169 cells by reverse transcription-quantitative PCR. The results confirm that the TLR transcripts from CRL2169 and HTB-9 cells used in this paper correspond to previously published results.

Article Snippet: The grade 2 male urothelial carcinoma cell line, HTB-9 (also called line 5637, ATCC-5637) and the grade 1 female urothelial carcinoma cell line, CRL-2169 (ATCC-SW 780) were grown in Lebovitz’s 15 (L-15) (ATCC-30-2008) medium supplemented with antibiotic-antimycotic (Sigma-A5955) and 10% FBS (Thermo-A3160401) at 37°C to roughly 80% confluency.

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction