Journal: PLOS One

Article Title: Integrated light and electron microscopy workflow for morphological, molecular and ultrastructural analysis of spheroids

doi: 10.1371/journal.pone.0342659

Figure Lengend Snippet: Representative haematoxylin–eosin (HE)-stained sections of SV-HUC-1 and T24 spheroids prepared by cryosectioning ( A-F ) and paraffin embedding (G-L) . A necrotic core (indicated by asterisks, A-B , D-E , G-H , J-K ) is visible in most cross sections, except those obtained from peripheral regions. Scale bars: 500 µm (A, D, G, J), 100 µm (B, E, H, K), 50 µm (C, F, I, L).

Article Snippet: Normal human urothelial SV-HUC-1 cells (CRL-9520) and muscle-invasive human bladder cancer urothelial T24 cells (HTB-4) were purchased from ATCC (Manassas, VA, United States) and cultured in a 1:1 mixture of A-DMEM medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) and F12 (Sigma-Aldrich, St. Louis, MO, United States), supplemented with 5% fetal bovine serum (Invitrogen, Carlsbad, CA, United States) and 4 mM GlutaMAX (Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Staining

Journal: PLOS One

Article Title: Integrated light and electron microscopy workflow for morphological, molecular and ultrastructural analysis of spheroids

doi: 10.1371/journal.pone.0342659

Figure Lengend Snippet: Representative images of paraffin sections show E-cadherin (green) in the plasma membrane in cells of SV-HUC-1 spheroids ( A-a3 ) and N-cadherin (red) in the plasma membrane of T24 spheroids ( B-b3 ). Some SV-HUC-1 cells are also positive for N-cadherin (arrows, a2 and a3 ). Yellow insets (in A and B) are magnified (a1-a3 and b1-b3) and display individual and merged channels of E-cadherin, N-cadherin, and DAPI-stained nuclei. Scale bars: 100 µm (A, B) , 20 µm (a1-a3, b1-b3).

Article Snippet: Normal human urothelial SV-HUC-1 cells (CRL-9520) and muscle-invasive human bladder cancer urothelial T24 cells (HTB-4) were purchased from ATCC (Manassas, VA, United States) and cultured in a 1:1 mixture of A-DMEM medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) and F12 (Sigma-Aldrich, St. Louis, MO, United States), supplemented with 5% fetal bovine serum (Invitrogen, Carlsbad, CA, United States) and 4 mM GlutaMAX (Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Clinical Proteomics, Membrane, Staining

Journal: PLOS One

Article Title: Integrated light and electron microscopy workflow for morphological, molecular and ultrastructural analysis of spheroids

doi: 10.1371/journal.pone.0342659

Figure Lengend Snippet: SV-HUC-1 (A) and T24 cells (C) form spheroids with a spherical morphology. (B) SV-HUC-1 cells are tightly attached to each other (arrows), and microvilli are seen on their surface. (D) T24 cells are loosely attached, displaying wider intercellular spaces (arrowheads) and have fewer microvilli. Scale bars: 100 µm (A, C) , 10 µm (B, D) .

Article Snippet: Normal human urothelial SV-HUC-1 cells (CRL-9520) and muscle-invasive human bladder cancer urothelial T24 cells (HTB-4) were purchased from ATCC (Manassas, VA, United States) and cultured in a 1:1 mixture of A-DMEM medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) and F12 (Sigma-Aldrich, St. Louis, MO, United States), supplemented with 5% fetal bovine serum (Invitrogen, Carlsbad, CA, United States) and 4 mM GlutaMAX (Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques:

Journal: PLOS One

Article Title: Integrated light and electron microscopy workflow for morphological, molecular and ultrastructural analysis of spheroids

doi: 10.1371/journal.pone.0342659

Figure Lengend Snippet: Outermost cells in SV-HUC-1 spheroids display cuboidal morphology (A) , whereas T24 cells are more elongated (B) . The presence of cell junctions in the outermost cell layer of SV-HUC-1 (C) and T24 spheroid (D) (boxed regions) indicates a tight cellular network of the outermost cell layer. The central necrotic zone (asterisks) is filled with necrotic cells in SV-HUC-1 and T24 spheroids (E, F) , as clearly demonstrated on semithin sections (G, H) , prepared before ultrathin sectioning. Scale bars: 100 µm (G-H) , 10 µm (A-B) , 5 µm (E-F) , 1 µm (C-D) .

Article Snippet: Normal human urothelial SV-HUC-1 cells (CRL-9520) and muscle-invasive human bladder cancer urothelial T24 cells (HTB-4) were purchased from ATCC (Manassas, VA, United States) and cultured in a 1:1 mixture of A-DMEM medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) and F12 (Sigma-Aldrich, St. Louis, MO, United States), supplemented with 5% fetal bovine serum (Invitrogen, Carlsbad, CA, United States) and 4 mM GlutaMAX (Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques:

Journal: Cancer Genomics & Proteomics

Article Title: Proteomic Signatures of Cellular Reprogramming in Bladder Cancer: Insights into the Acquisition of Cancer Stem-like States and Phenotypic Plasticity

doi: 10.21873/cgp.20578

Figure Lengend Snippet: Heatmap analysis of global and functional protein expression during bladder cancer cell reprogramming. (A) Whole proteome clustering reveals distinct segregation of reprogrammed bladder cancer cell passages and partial convergence toward SV-HUC-1. (B) Proteins associated with cell adhesion exhibit partial restoration of epithelial-like profile. (C) Proteins in cellular developmental process show progressive modulation in early cellular reprogramming. (D) Cell motility-related proteins exhibit reduced expression in later reprogramming passages, suggesting a shift away from a migratory phenotype. (E) Proteins related to epithelial cell differentiation in rep HTB-4 cells demonstrate increased similarity to SV-HUC-1, highlighting the re-establishment of epithelial characteristics. Color scale represents Z-score–normalized abundance values (green=low; red=high).

Article Snippet: HTB-4 (ATCC HTB-4; RRID: CVCL_0554) and SV-HUC-1 (ATCC CRL-9520; RRID: CVCL_3798) cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Functional Assay, Expressing, Cell Differentiation

Journal: Cancer Genomics & Proteomics

Article Title: Proteomic Signatures of Cellular Reprogramming in Bladder Cancer: Insights into the Acquisition of Cancer Stem-like States and Phenotypic Plasticity

doi: 10.21873/cgp.20578

Figure Lengend Snippet: Comparative proteomic analysis of reprogrammed bladder cancer cells and normal uroepithelial SV-HUC-1 cells. (A) Venn diagrams showing the total numbers of regulated, upregulated, and downregulated proteins shared or uniquely detected at passages p1, p4, and p7. (B) Protein-protein interaction (PPI) network of proteins commonly upregulated in all three passages (p1, p4, p7) in comparison to the normal uroepithelial cell line SVHUC- 1. Nodes represent proteins and edges show experimentally validated interactions. Enriched pathways are indicated in different colors. (C) PPI network of commonly downregulated proteins across rep HTB-4 p1, p4, p7. (D) Gene ontology (GO) analysis showed enrichment of commonly upregulated proteins in rep HTB-4 p1, p4, and p7 cells, involved in biological processes, molecular function, and cellular components [fold change >2 and false discovery rate (FDR)<0.01]. Significant overrepresentation of processes related to biosynthesis, peptide metabolic pathways, RNA binding, and cadherin binding was detected. (E) Gene ontology (GO) analysis showed enrichment of commonly downregulated proteins in rep HTB-4 p1, p4, and p7 cells, involved in biological processes, molecular function, and cellular components [fold change >2 and false discovery rate (FDR) <0.01]. Reductions in pathways linked to RNA processing, the catenin complex, and the ribonucleoprotein complex were detected.

Article Snippet: HTB-4 (ATCC HTB-4; RRID: CVCL_0554) and SV-HUC-1 (ATCC CRL-9520; RRID: CVCL_3798) cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Comparison, RNA Binding Assay, Binding Assay