Journal: Molecules

Article Title: Exploring EGFR, Nectin-4, and TROP-2 as Therapeutic Targets for Bladder Cancer Photoimmunotherapy

doi: 10.3390/molecules30244802

Figure Lengend Snippet: ( a ) Western blot analysis of EGFR, Nectin-4, and TROP-2 expression in the BC cell lines RT4 and RT112 and in the antigen-negative control cell line CHO. ( b ) Cell binding of the antibodies and conjugates to the target cells as demonstrated by flow cytometry. Binding of the isotype control antibody is shown in gray.

Article Snippet: RT4 cells were cultivated in EMEM medium (Cytion, Eppelheim, Germany), RT112 cells in RPMI1640 medium (Gibco), and CHO cells in F-12 Nutrient Mixture Medium (Gibco) at 37 °C and 5% CO 2 .

Techniques: Western Blot, Expressing, Negative Control, Binding Assay, Flow Cytometry, Control

Journal: Molecules

Article Title: Exploring EGFR, Nectin-4, and TROP-2 as Therapeutic Targets for Bladder Cancer Photoimmunotherapy

doi: 10.3390/molecules30244802

Figure Lengend Snippet: Cytotoxicity of the conjugates Cmb T120C/D265C -WB692-CB2, Enf T120C/D265C -WB692-CB2, and Sac T120C/D265C -WB692-CB2 after irradiation with different light doses in RT4 and RT112 cells. CHO cells served as antigen-negative controls. Mean values ± SD of three independent biological experiments. Statistical analyses were performed using unpaired, parametric Student’s t -tests with Welch’s correction (* p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001).

Article Snippet: RT4 cells were cultivated in EMEM medium (Cytion, Eppelheim, Germany), RT112 cells in RPMI1640 medium (Gibco), and CHO cells in F-12 Nutrient Mixture Medium (Gibco) at 37 °C and 5% CO 2 .

Techniques: Irradiation

Journal: Molecules

Article Title: Exploring EGFR, Nectin-4, and TROP-2 as Therapeutic Targets for Bladder Cancer Photoimmunotherapy

doi: 10.3390/molecules30244802

Figure Lengend Snippet: Cytotoxicity of the conjugates Cmb T120C/D265C -WB692-CB2, Enf T120C/D265C -WB692-CB2 and Sac 120T/265C -WB692-CB2 alone and in combination after irradiation with a light dose of 64 J/cm 2 in RT4 and RT112 cells. Control samples were treated with the uncoupled antibodies or the free dye. Mean values ± SD of three independent biological experiments. Statistical analyses were performed using unpaired, parametric Student’s t -tests with Welch’s correction (* p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001).

Article Snippet: RT4 cells were cultivated in EMEM medium (Cytion, Eppelheim, Germany), RT112 cells in RPMI1640 medium (Gibco), and CHO cells in F-12 Nutrient Mixture Medium (Gibco) at 37 °C and 5% CO 2 .

Techniques: Irradiation, Control

Journal: International Journal of Molecular Sciences

Article Title: Tyro3 Targeting as a Radiosensitizing Strategy in Bladder Cancer through Cell Cycle Dysregulation

doi: 10.3390/ijms23158671

Figure Lengend Snippet: Impact of the modulation of TYRO3 expression on the radiosensitivity of bladder cancer cell lines. ( a – c ) Representative clonogenic survival curves of RT112 and 5637 cell lines ( upper panel). 48 h after transfection with siLUC (control, red), siTYRO3#4 (blue), siTYRO3#801 (green) or diluted concentration of siRNA#1 (black) cells were exposed to increased doses of gamma-rays. In each case, downregulation was confirmed by western blot at 48 h after transfection. The corresponding D 10 values were calculated after fitting the experimental data to the classical linear-quadratic equation ( lower panel); ( d ) Representative survival curves of UM-UC-3 control (empty plasmid, red) and TYRO3 over-expressed cell lines (TYRO3 encoding- plasmid, purple) ( upper panel). The upregulation was confirmed by western blot. The corresponding D 10 values were calculated after fitting the experimental data to the classical linear-quadratic equation ( lower panel); ( e ) Phosphorylated TYRO3 shown in western blot ( lower panel) after immunoprecipitation of the cell extracts with phospho-Tyrosine antibodies and its relative expression in 5637 cells quantified using Image-J software and normalized to that of total TYRO3 ( upper panel). Data represents mean ± SD of 3 independent experiments. Unpaired t-test analysis: * p < 0.05; ** p < 0.005, ns: not significant.

Article Snippet: The human BCa-derived cell lines 5637, RT112, UM-UC-5, UM-UC-9, VM-CUB-1, were obtained from DSMZ (Heidelberg, Germany), and UM-UC-3 from ATCC.

Techniques: Expressing, Transfection, Control, Concentration Assay, Western Blot, Plasmid Preparation, Immunoprecipitation, Software

Journal: International Journal of Molecular Sciences

Article Title: Tyro3 Targeting as a Radiosensitizing Strategy in Bladder Cancer through Cell Cycle Dysregulation

doi: 10.3390/ijms23158671

Figure Lengend Snippet: TYRO3 modulation and its impact on Ionizing Radiation-Induced Foci and DNA damage. γH2AX foci visualized after 24 h of 2 Gy irradiation in TYRO3 downregulated RT112 ( a ) or 5637 ( c ) cells (scale bar 5 microns); Quantification of cells containing more than 10 γH2AX foci at 24 h after 2 Gy of irradiation in the downregulated RT112 ( b ) and 5637 ( d ) cell lines; ( e ) γH2AX foci visualized after 24 h of 2 Gy irradiation in TYRO3 overexpressing UM-UC-3 cell line (Scale bar 5 microns); ( f ) Quantification of cells containing more than 10 γH2AX foci at 30 min and 24 h after 2 Gy of irradiation in TYRO3 overexpressing cell lines. The data shown above is from three different experiments and error bars represent the SD. Unpaired t -test analysis: * p < 0.05; ** p < 0.005; *** p < 0.0005; **** p < 0.0005, ns: non-significant. Representative images of the alkaline Comet assay performed on TYRO3 downregulated RT112 ( g ) and 5637 ( h ) cells and the resulting Olive tail moments analysis in TYRO3 downregulated RT112 ( i ) and 5637 ( j ) cells irradiated at 6 Gy. The data shown here is from three independent experiments analyzing 200 nucleus per condition per experiment, horizontal bars represent the median values. Kruskal-Wallis nonparametric tests with Multiple comparisons were used: * p < 0.05; ** p < 0.005, ns: non-significant; ( k ) western blot of DNA damage repair proteins; TYRO3 was downregulated, irradiated at 6 Gy and the lysates were prepared and analyzed 0.5–24 h after.

Article Snippet: The human BCa-derived cell lines 5637, RT112, UM-UC-5, UM-UC-9, VM-CUB-1, were obtained from DSMZ (Heidelberg, Germany), and UM-UC-3 from ATCC.

Techniques: Irradiation, Alkaline Single Cell Gel Electrophoresis, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Tyro3 Targeting as a Radiosensitizing Strategy in Bladder Cancer through Cell Cycle Dysregulation

doi: 10.3390/ijms23158671

Figure Lengend Snippet: TYRO3 modulation and its impact on DNA damage response pathways. ( a ) Volcano plot showing the fold change (Log2 Ratio) versus negative log of the p-value of differentially expressed genes after Nanostring analysis between BCa (RT112 and 5637) 6 Gy irradiated cells versus BCa 6 Gy irradiated cells after complete TYRO3 knock-down (siTYRO3#4 and siTYRO3#801). Significant: p -value < 0.05 ( b ) Log2 ratio of the significantly expressed genes ( p < 0.05) after Nanostring analysis between the two groups. The name of the up- or downregulated genes are listed on the left. On the right are the corresponding Nanostring gene annotations. ( c ) Protein-protein association network of the up and downregulated genes assessed using the STRING database.

Article Snippet: The human BCa-derived cell lines 5637, RT112, UM-UC-5, UM-UC-9, VM-CUB-1, were obtained from DSMZ (Heidelberg, Germany), and UM-UC-3 from ATCC.

Techniques: Irradiation, Knockdown

Journal: International Journal of Molecular Sciences

Article Title: Tyro3 Targeting as a Radiosensitizing Strategy in Bladder Cancer through Cell Cycle Dysregulation

doi: 10.3390/ijms23158671

Figure Lengend Snippet: TYRO3 downregulation affects cell cycle following irradiation. Analysis of the cell cycle distribution 24 h after 6 Gy irradiation on TYRO3-downregulated RT112 ( a ) and 5637 ( b ) cells. Comparison of percentage (%) of cells in G2/M in RT112 ( c ) and 5637 ( d ) cells. Data shown is from three different experiments and error bars represent the SD. Unpaired t-test analysis: * p < 0.05; ** p < 0.005; *** p < 0.0005, ns: non-significant. ( e ) western blot of cell cycle proteins 30 min and up to 24 h after irradiation.

Article Snippet: The human BCa-derived cell lines 5637, RT112, UM-UC-5, UM-UC-9, VM-CUB-1, were obtained from DSMZ (Heidelberg, Germany), and UM-UC-3 from ATCC.

Techniques: Irradiation, Comparison, Western Blot

Journal: Oncogenesis

Article Title: Adaptation of redox metabolism in drug-tolerant persister cells is a vulnerability to prevent relapse in pancreatic cancer

doi: 10.1038/s41389-025-00591-0

Figure Lengend Snippet: A Schematic of experimental protocol. Orthotopic syngeneic allografts were generated by intraperitoneal injection of one million of the murine KPCluc2 cells into 5-week-old female C57BL/6 immunocompetent mice. Tumoral growth was followed by bioluminescence using a Photon Imager device (Biospace Lab). Twelve days post-grafting, tumor-bearing mice were randomized to two treatment cohorts, vehicle control (PBS) and gemcitabine during one month. Mice were sacrificed according to the ethical limit point . B , C Longitudinal in vivo bioluminescence imaging analysis of KPCluc2 cell-grafted mice, untreated (Control) or treated with gemcitabine starting 12 days after implantation. B Bioluminescence images. C Bioluminescent signal quantification in which each line, solid or dotted, represents one mouse (RLU = relative light unit). Tumors in mice 2, 3, 4, 5, 7, and 8 responded to gemcitabine treatment but did not completely regress (Gem group); their numbers are shown in pink. In contrast, tumors in mice 1, 6, 9, 10, 11, and 12 regressed completely as evidenced by the absence or very low level of bioluminescence at at least one stage of treatment effect analysis (Gem-CR group); their numbers are shown in purple. The bioluminescent signal is expressed in photons per second per square centimeter per steradian (ph/s/cm 2 /sr). D Mitochondrial mass, MMP, Mitochondrial O2.- and total ROS level were measured in tumors as in Fig. . Median fluorescence intensity (MFI) is shown relative to unlabeled cells. Each dot corresponds to one mouse, and the bars show the mean ± SD. E Total ATP level was measured using the cell viability assay (Cell-Titer Glo Kit). Unpaired Student’s T -test was used for statistical analyzes comparing each group with that of the untreated. *, **, *** and **** correspond to p < 0.05, 0.01, 0.001, and 0.0001, respectively; ns non-significant difference.

Article Snippet: One million KPCluc2 cells were IP injected into 5- to 6-week-old C57BL/6 female mice (immunocompetent strain, SOPF health status, Charles River, France).

Techniques: Generated, Injection, Control, In Vivo, Imaging, Fluorescence, Viability Assay