human normal bladder epithelial cell line sv huc  (ATCC)

Journal: Nature Communications

Article Title: Multi-omics analysis identifies a microbiota–bile acid–TLR signaling axis driving bladder injury in interstitial cystitis

doi: 10.1038/s41467-025-68060-1

Figure Lengend Snippet: A Differentially expressed genes (DEGs) in full-thickness bladder tissues from HIC patients and controls identified via RNA sequencing (|log₂FC|≥2; adjusted p < 0.05, two-sided Wald test using DESeq2). Gene Ontology (GO) ( B ) and KEGG ( C ) enrichment analyses of DEGs (two-sided hypergeometric test with FDR correction). D UMAP visualization of single-cell transcriptomic landscape of urothelium, with urothelial cells (UCs) identified by KRT19 and UPK1A expression. GO ( E ) and KEGG ( F ) analyses of upregulated DEGs in UCs (two-sided hypergeometric test with FDR correction). GO ( G ) and KEGG ( H ) analyses of downregulated DEGs in UCs (two-sided hypergeometric test with FDR correction). I , J Gene Set Enrichment Analysis (GSEA) of altered pathways in UCs (two-sided permutation test with 1000 permutations, with multiple testing corrected using the FDR). The exact p value for TLR signaling of UCs in panel I is 0.0009. K Expression of Toll-like receptor (TLR) subtypes in UCs from HIC bladders. mRNA ( L ) and protein ( M ) expression levels of TLR3 in isolated urothelium from control and HIC patients ( n = 7 control vs. 10 HIC; data show median (IQR); two-sided Mann–Whitney U test; ns not significant; Bar for panel M: 1 cm). N Immunostaining showing the distribution of TLR3 protein in the urothelium of patients with HIC ( n = 7 control vs. 10 HIC; one section and field per patient; Bar: 100 μm). IHC immunohistochemistry, HL Hunner lesions, U urothelium, LP lamina propria (blue line), M muscularis. Source data are provided as a Source Data file.

Article Snippet: Primary bladder urothelial cells (ATCC, Cat. PCS-420-010) were cultured in bladder epithelial cell basal medium (ATCC, Cat. PCS-420-032) supplemented with bladder epithelial growth kit (ATCC, Cat. PCS-420-042) at 37 °C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: RNA Sequencing, Expressing, Isolation, Control, MANN-WHITNEY, Immunostaining, Immunohistochemistry

Journal: Nature Communications

Article Title: Multi-omics analysis identifies a microbiota–bile acid–TLR signaling axis driving bladder injury in interstitial cystitis

doi: 10.1038/s41467-025-68060-1

Figure Lengend Snippet: A Fecal microbiota transplantation (FMT) in antibiotic-pretreated mice (Abx-mice), using stool samples from healthy controls (Group A, No. of controls = 5) and HIC patients (Group B, No. of patients = 5). B Bladder weight comparison post-FMT. C Assessment of voiding behavior in FMT-recipient mice (each mouse measured once). D Evaluation of mechanical pain threshold following FMT. Comparisons: (1) Control vs. Autoimmune cystitis (AC); (2) Control vs. Control + A; (3) Control vs. Control + B; (4) AC vs. AC + A; (5) AC vs. AC + B. E Representative histological analysis of bladder tissues after FMT (one section and field per mouse). F Quantification of urinary TCDCA and TUDCA in recipient mice. For panels ( B – F ): group sizes are n = 7 for Control + A and n = 8 for all other groups; data are presented as median (IQR); statistical analysis was performed using the Kruskal–Wallis test; ns not significant. IHC immunohistochemistry, Red arrow: urothelial thinning, detachment, and exposure, Blue line: LP lamina propria, U urothelium, M muscularis. Source data are provided as a Source Data file.

Article Snippet: Primary bladder urothelial cells (ATCC, Cat. PCS-420-010) were cultured in bladder epithelial cell basal medium (ATCC, Cat. PCS-420-032) supplemented with bladder epithelial growth kit (ATCC, Cat. PCS-420-042) at 37 °C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Transplantation Assay, Comparison, Control, Immunohistochemistry

Journal: Nature Communications

Article Title: Multi-omics analysis identifies a microbiota–bile acid–TLR signaling axis driving bladder injury in interstitial cystitis

doi: 10.1038/s41467-025-68060-1

Figure Lengend Snippet: A Experimental workflow of E. avium colonization in antibiotic-pretreated mice (Abx-mice, n = 6 per group). Created in BioRender. Chen, J. (2025) https://BioRender.com/3lccwo1 . B Increased abundance of E. avium confirmed by metagenomic sequencing ( n = 6 per group). C Evaluation of voiding behavior following E. avium transplantation ( n = 6 per group; one measurement per mouse). D Measurement of mechanical pain threshold post-transplantation ( n = 6 per group). E Representative histological analysis of bladder tissues after E. avium transplantation ( n = 6 per group; one section and field per mouse). F , G Quantification of blood and urinary bile acids using comprehensive targeted bile acid profiling ( n = 6 per group). H Cell viability of human urothelial cells (HUCs) following TCDCA or TUDCA treatment ( n = 3 independent experiments). I Expression of ZO-1, TNF-α, and TLR3 following TCDCA (200 µM) exposure ( n = 3 independent experiments). J Experimental workflow of intravesical instillation of TCDCA in rats ( n = 5 per group). Created in BioRender. Chen, J. (2025) https://BioRender.com/3lccwo1 . K Evaluation of voiding function after TCDCA instillation ( n = 5 per group; each rate measured once). L Assessment of mechanical pain threshold following TCDCA exposure ( n = 5 per group). M Representative histology of bladder tissues post-instillation ( n = 5 per group; one section per rat, one field quantified per section). For panels ( B – H , K , L ): data are presented as median (IQR). Statistical analysis was performed using the two-sided Mann–Whitney U test. IHC immunohistochemistry, Red arrow: urothelial thinning, detachment, and exposure, Blue line: LP lamina propria, U urothelium, M muscularis. Source data are provided as a Source Data file.

Article Snippet: Primary bladder urothelial cells (ATCC, Cat. PCS-420-010) were cultured in bladder epithelial cell basal medium (ATCC, Cat. PCS-420-032) supplemented with bladder epithelial growth kit (ATCC, Cat. PCS-420-042) at 37 °C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Sequencing, Transplantation Assay, Expressing, MANN-WHITNEY, Immunohistochemistry

Journal: Nature Communications

Article Title: Multi-omics analysis identifies a microbiota–bile acid–TLR signaling axis driving bladder injury in interstitial cystitis

doi: 10.1038/s41467-025-68060-1

Figure Lengend Snippet: A , B Expression of tight junction protein ZO-1 and inflammatory marker TNF-α after TLR3 intervention in TCDCA-pretreated human urothelial cells (HUCs) ( n = 3 independent experiments). C Transepithelial resistance (TER) changes following TLR3 intervention in TCDCA-pretreated HUCs ( n = 3 independent experiments). D , E Expression of ZO-1 and TNF-α after pentosan polysulfate sodium (PPS) administration in TCDCA-pretreated HUCs ( n = 3 independent experiments). F TER changes following PPS intervention in TCDCA-pretreated HUCs ( n = 3 independent experiments). G Experimental workflow showing TLR3 inhibitor and PPS administration to rats following intravesical TCDCA instillation ( n = 5 per group). Created in BioRender. Chen, J. (2025) https://BioRender.com/3lccwo1 . H Assessment of voiding function (one measurement per rat) and pain threshold after TLR3 inhibition ( n = 5 per group). I Representative histological images of bladder tissues post-TLR3 inhibition ( n = 5 per group; one section and field per rat). For panels ( B , C , E , F , H ): data are presented as median (IQR). Statistical analysis was performed using the Kruskal–Wallis test. ns not significant, IHC immunohistochemistry, Red arrow: urothelial thinning, detachment, and exposure, Blue line: LP lamina propria, U urothelium, M muscularis. Source data are provided as a Source Data file.

Article Snippet: Primary bladder urothelial cells (ATCC, Cat. PCS-420-010) were cultured in bladder epithelial cell basal medium (ATCC, Cat. PCS-420-032) supplemented with bladder epithelial growth kit (ATCC, Cat. PCS-420-042) at 37 °C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Expressing, Marker, Inhibition, Immunohistochemistry

Journal: Biochemistry and Biophysics Reports

Article Title: AEBP1-GLI1 pathway attenuates the FACT complex dependency of bladder cancer cell survival

doi: 10.1016/j.bbrep.2025.102101

Figure Lengend Snippet: AEBP1 expression in bladder cancer cell lines. A, Gene chip analysis for AEBP1 expression in normal and cancer bladders using TNMplot software ( https://tnmplot.com/analysis/ ). B, Kaplan-Meier analysis of overall survival of patients with bladder cancer using UCSCxena software ( https://xena.ucsc.edu/ ). High expression of AEBP1 negatively impacts overall survival. C, AEBP1 expression in normal human bladder epithelial cells (HBEC) and bladder cancer cell lines. D, Cell proliferation of AEBP1 -high (5637 and KU1919) and AEBP1 -low (JMSU1) cells. Data are shown as mean + SD from six biological replicates. P ‐values obtained by Student's t‐tests. P < 0.001 was set as statistically significant. Double asterisk, P < 0.001. E, Analyses of AEBP1 knockdown. 72 h after transfection with siRNAs for AEBP1 (siAEBP1-1 and siAEBP1-4) or their control (siControl), cells were harvested for the immunoblot analyses. F, Ratios of cell numbers 72 h after transfection of siControl (C), siAEBP1-1 (1), or siAEBP1-4 (4). Data are shown as mean + SD from four or three biological replicates. P ‐values obtained by Student's t‐tests. P < 0.001 was considered as statistically significant. Double asterisk, P < 0.001. NS, not significant.

Article Snippet: Primary cultures of normal human bladder epithelial cells (HBEC) were obtained from KURABO and maintained in culture medium according to the manufacturer's protocols.

Techniques: Expressing, Software, Knockdown, Transfection, Control, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: AEBP1-GLI1 pathway attenuates the FACT complex dependency of bladder cancer cell survival

doi: 10.1016/j.bbrep.2025.102101

Figure Lengend Snippet: FACT complex independency of AEBP1 -high cancer cells. A, Volcano plot of RNA-seq data. 72 h after transfection with either siControl or siAEBP1-1, 5637 cells were harvested for RNA extraction followed by RNA-seq analyses. Blue and red dots show down- and upregulated genes by AEBP1 knockdown, respectively. RNA-seq data was registered in the GEO database (NCBI) under the accession number GSE288105 . B, Expression of SSRP1 and SUPT16H proteins in normal human bladder epithelial cells (HBEC) and bladder cancer cell lines. C, Induction of the cleaved form of PARP (cPARP), Caspase3 (cCaspase3), and the phosphorylated form of histone H2A (γ-H2AX). Cells were harvested for the immunoblot analyses 72 h after siRNA transfection. Double knockdown of SSRP1 and SUPT16H enhances the induction of cPARP, cCaspase3, and γ-H2AX in JMSU1 but not 5637 cells. In 5637 cells, AEBP1 knockdown induces these markers more than SSRP1 and SUPT16H double knockdown. Note that consistent with RNA-seq data, AEBP1 knockdown reduced the protein levels of SSRP1 and SUPT16H in 5637 cells, but not in JMSU1 cells. The signal intensity of SUPT16H and SSRP1 was semi-quantified using ImageJ software.

Article Snippet: Primary cultures of normal human bladder epithelial cells (HBEC) were obtained from KURABO and maintained in culture medium according to the manufacturer's protocols.

Techniques: RNA Sequencing, Transfection, RNA Extraction, Knockdown, Expressing, Western Blot, Software

Journal: Biochemistry and Biophysics Reports

Article Title: AEBP1-GLI1 pathway attenuates the FACT complex dependency of bladder cancer cell survival

doi: 10.1016/j.bbrep.2025.102101

Figure Lengend Snippet: GLI1 defines the FACT complex independency in AEBP1 -high cancer cells. A, Correlated expression of GLI1 and GLI2 with AEBP1 in bladder cancer. TCGA data (n = 436) was analyzed using USCSxena software ( https://xena.ucsc.edu/ ). B, Expression of GLI1 and GLI2 proteins in normal human bladder epithelial cells (HBEC) and bladder cancer cell lines. C, AEBP1 knockdown reduces GLI1 protein in 5637 and KU1919 cells. D, GANT61-induced cPARP, cCaspase3, and γ-H2AX were not suppressed by AEBP1 overexpression. Cells were treated with GANT61 (10 and 12 μM) or its vehicle DMSO (0 μM) for 72 h. For the overexpression experiments, 5637 cells were transduced with either AEBP1 expressing lentivirus (Lenti-AEBP1) or its empty control lentivirus (Lenti-control) and maintained under blasticidin. Immunoblot analyses detected the expression of endogenous and virally transduced AEBP1 via short or long exposure. E, Scheme of AEBP1 role in cancer cell survival. In AEBP1 -high cancer cells, GLI1 bypasses the FACT complex to maintain the cell survival, resulting in limited FACT complex dependency of cancer cell survival.

Article Snippet: Primary cultures of normal human bladder epithelial cells (HBEC) were obtained from KURABO and maintained in culture medium according to the manufacturer's protocols.

Techniques: Expressing, Software, Knockdown, Over Expression, Transduction, Control, Western Blot