htb 9  (ATCC)

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

Journal: eBioMedicine

Article Title: Somatic structural variants drive upper tract urothelial carcinoma muscle invasiveness via activation of TPX2 transcription

doi: 10.1016/j.ebiom.2026.106182

Figure Lengend Snippet: An upstream structural variant interval enhances TPX2 expression and promotes invasive phenotypes in urothelial cancer cells. A. Schematic representation of somatic structural variants (SSVs) surrounding the TPX2 locus. A recurrent duplication event localised to the chr20:31000852–31001508 interval upstream of TPX2 was identified. This region contains eight tandem repeats of a 171-bp sequence. Two unique sgRNAs were designed to delete a 683-bp fragment spanning this interval. B. PCR and gel electrophoresis validation of CRISPR ribonucleoprotein (RNP)-mediated deletion of the TPX2 upstream interval in 5637 cells. The main band corresponding to the intact region (1025 bp) was markedly attenuated in TPX2-upstream knockout cells. (C, D). Quantitative RT-PCR (C) and Western blot (D) analyses showing that deletion of the chr20:31000852–31001508 interval significantly reduced TPX2 mRNA and protein expression in 5637 cells. (E, F) Quantitative RT-PCR (E) and Western blot (F) analyses demonstrating that transfection of a PCR-derived fragment corresponding to the TPX2 upstream interval increased TPX2 expression compared with control cells. G. Schematic of the dual-luciferase reporter construct in which the TPX2 promoter region (2331 bp, including exon 1) was cloned upstream of the Renilla luciferase (Rluc) gene in the psiCHECK-2 vector. H. Dual-luciferase reporter assay showing that co-transfection of the TPX2 promoter reporter with PCR-derived fragments from the chr20:31000852–31001508 interval significantly increased Rluc activity compared with co-transfection with a random 1-kb DNA fragment, indicating enhancer-like activity of this interval. (I, J) Western blot validation of TPX2 knockout (KO) (I) and TPX2 overexpression (OE) (J) in 5637 cells. (K, L) Wound-healing (K) and transwell migration and invasion assays (L) demonstrating that TPX2 knockout significantly impaired cell motility and invasive capacity. Scale bar = 1 mm in K, scale bar = 200 um in L. (M, N) Wound-healing (M) and transwell migration and invasion assays (N) showing that TPX2 overexpression markedly enhanced migratory and invasive properties of 5637 cells. Scale bar = 1 mm in M, scale bar = 200 um in N. Data are presented as mean ±SD (n = 3). Two-sided Mann-Whitney U test was used for comparisons between two groups. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001). O. KEGG pathway enrichment analysis of genes downregulated upon TPX2 knockout (TPX2_KO vs. NC_KO). P. Gene set enrichment analysis (GSEA) of transcriptomic changes induced by amplification of the TPX2 upstream interval (Ups_OE vs. NC_OE). Pathways related to EMT, cell adhesion, and extracellular matrix remodelling were significantly enriched.

Article Snippet: The human bladder cancer-derived 5637 cells (ATCC) were cultured in RPMI 1640 culture medium supplemented with 10% foetal bovine serum (FBS) (Gibco) and 1% penicillin/streptomycin (Invitrogen) at 37 °C with 5% CO2 and maximum humidity.

Techniques: Variant Assay, Expressing, Sequencing, Nucleic Acid Electrophoresis, Biomarker Discovery, CRISPR, Knock-Out, Quantitative RT-PCR, Western Blot, Transfection, Derivative Assay, Control, Luciferase, Construct, Clone Assay, Plasmid Preparation, Reporter Assay, Cotransfection, Activity Assay, Over Expression, Migration, MANN-WHITNEY, Amplification