mouse renal tubular epithelial cells mrtecs  (ATCC)

Journal: Cell Biology and Toxicology

Article Title: Early endolysosomal dysfunction is a contributing factor to gadolinium-based contrast agent mouse renal proximal tubule epithelial cell injury

doi: 10.1007/s10565-025-10014-w

Figure Lengend Snippet: Intracellular accumulation of linear and macrocyclic gadolinium-based contrast agents. a . Chemical structures, chemical names, and generic and brand names of the linear and macrocyclic gadolinium-based contrast agents used in this study. b . Gadolinium concentrations in MRPTEpiCs at 1, 4, 24, and 48 h GBCA exposure, as assayed by ICP-MS. Gadolinium levels were obtained from 1 × 10 6 cells, with each value representing the mean of triplicate experiments ± SEM ( n = 3 per group). The p -values are denoted numerically, from Untreated, by One-way ANOVA, Tukey honesty significant difference post-hoc testing

Article Snippet: Mouse renal proximal tubular epithelial cells (MRPTEpiCs) (ScienCell Research Laboratories, #M4100-57) were cultured according to recommended protocols in complete epithelial cell medium-animal (c/EpiCM-a) (ScienCell Research Laboratories, #4131) and maintained at 37oC and 5% CO 2 .

Techniques:

Journal: Cell Biology and Toxicology

Article Title: Early endolysosomal dysfunction is a contributing factor to gadolinium-based contrast agent mouse renal proximal tubule epithelial cell injury

doi: 10.1007/s10565-025-10014-w

Figure Lengend Snippet: GBCA-induced lysosomal enlargement and injury. a . Lysosomal-associated membrane protein 1 (LAMP-1) (green) expression and distribution in untreated and GBCA exposed cells. Nuclei counterstained with DAPI. Higher magnification of the region of interest is denoted by the dashed box and displayed adjacent to the respective image. Leica TCS SP8. Bars = 10 µm b . Quantification of LAMP-1 granule diameter in untreated and treated cells following 4 h and 24 h exposures. Results are represented as mean ± SEM from ≥ 50 cells ( n = 3). The p -values are denoted numerically and determined by Two-way ANOVA, Tukey honesty significant difference post-hoc testing. c . Representative images of galectin-3 translocation assay in GBCA-exposed MRPTEpiCs at the indicated time points. Higher magnification of the merged region of interest is denoted by the dashed box and displayed adjacent to the respective image: Galectin-3 (Upper), LAMP-1 (Middle), Merge (Lower). Leica TCS SP8. Bars = 10 µm. d . Quantification of galectin-3 positive cells in untreated and treated MRPTEpiCs at 4 h and 24 h GBCA exposure. Results are represented as mean ± SEM from ≥ 75 cells ( n = 3). The p -values are denoted numerically and determined by Two-way ANOVA, Tukey honesty significant difference post-hoc testing. e . Transmission electron microscopy images of MRPTEpiCs following 4 and 24 h exposure highlighting vacuolization (magenta arrows). Nuclear fragmentation (black arrow). n ≥ 75 cells per group. Hitachi HT7000 TEM, AMT 16-megapixel digital camera. Bars = 2.5 µm

Article Snippet: Mouse renal proximal tubular epithelial cells (MRPTEpiCs) (ScienCell Research Laboratories, #M4100-57) were cultured according to recommended protocols in complete epithelial cell medium-animal (c/EpiCM-a) (ScienCell Research Laboratories, #4131) and maintained at 37oC and 5% CO 2 .

Techniques: Membrane, Expressing, Translocation Assay, Transmission Assay, Electron Microscopy

Journal: Cell Biology and Toxicology

Article Title: Early endolysosomal dysfunction is a contributing factor to gadolinium-based contrast agent mouse renal proximal tubule epithelial cell injury

doi: 10.1007/s10565-025-10014-w

Figure Lengend Snippet: Effect of GBCAs on cathepsin B processing and lysosomal membrane permeabilization. a . Representative western blot showing levels of CTSB in MRPTEpiCs following 4 h exposure. b-c . Quantitative analysis of relative proCTSB and mCTSB levels in untreated and exposed groups, respectively. d . Western blot displaying the expression levels of CTSB after 24 h linear or macrocyclic GBCA exposure. e–f . Quantification of relative protein levels of proCTSB and mCTSB following exposure. b-c-e–f. Data represented as mean ± SEM ( n = 4 per group). The p -values are denoted numerically, ns = not significant from Untreated, by One-way ANOVA, Tukey honesty significant difference post-hoc testing. g . Representative confocal images of CTSB and LAMP-1 staining patterns in GBCA-exposed MRPTEpiCs from at least three independent experiments. Inset dashed boxes indicate regions of interest highlighted by magnified adjacent images: CTSB (Upper), LAMP-1 (Middle), and Merge (Lower). Nuclei were counterstained with DAPI. Leica TCS SP8. Bars = 10 µm

Article Snippet: Mouse renal proximal tubular epithelial cells (MRPTEpiCs) (ScienCell Research Laboratories, #M4100-57) were cultured according to recommended protocols in complete epithelial cell medium-animal (c/EpiCM-a) (ScienCell Research Laboratories, #4131) and maintained at 37oC and 5% CO 2 .

Techniques: Membrane, Western Blot, Expressing, Staining

Journal: Cell Biology and Toxicology

Article Title: Early endolysosomal dysfunction is a contributing factor to gadolinium-based contrast agent mouse renal proximal tubule epithelial cell injury

doi: 10.1007/s10565-025-10014-w

Figure Lengend Snippet: Impact of GBCA exposure on cathepsin D processing and release from lysosomal lumen. a . Western blot showing levels of CTSD in 4 h exposed MRPTEpiCs. b-c . Quantification of relative proCTSD and mCTSD levels in untreated and exposed groups. d . Representative western blot displaying expression levels of CTSD following 24 h GBCA exposures. e–f . Quantitative analysis of relative protein levels of proCTSD and mCTSD after exposure. b-c - e–f . Results are represented as mean ± SEM ( n = 4 per group). The p -values are denoted numerically, ns = not significant, by One-way ANOVA, Tukey honesty significant difference post-hoc testing. g . Confocal images of GBCA-exposed MRPTEpiCs highlighting CTSD staining patterns relative to LAMP-1 staining from at least three independent experiments. Inset dashed boxes indicate regions of interest emphasized by magnified adjacent images: CTSD (Upper), LAMP-1 (Middle), and Merge (Lower). Nuclei were counterstained with DAPI. Leica TCS SP8. Bars = 10 µm

Article Snippet: Mouse renal proximal tubular epithelial cells (MRPTEpiCs) (ScienCell Research Laboratories, #M4100-57) were cultured according to recommended protocols in complete epithelial cell medium-animal (c/EpiCM-a) (ScienCell Research Laboratories, #4131) and maintained at 37oC and 5% CO 2 .

Techniques: Western Blot, Expressing, Staining

Journal: Cell Biology and Toxicology

Article Title: Early endolysosomal dysfunction is a contributing factor to gadolinium-based contrast agent mouse renal proximal tubule epithelial cell injury

doi: 10.1007/s10565-025-10014-w

Figure Lengend Snippet: Effect of lysosomal protease inhibition on the mitochondria structure and function in GBCA-treated MRPTEpiCs. a . Representative confocal images of labeled mitochondria following 24 h GBCA exposure plus co-treatment with the indicated lysosomal protease inhibitors. Live cells loaded with 250 nM MitoTracker Red-FM. Leica TCS SP8. Bars = 10 µm. Insets are processed, binary images produced in FIJI/Image J . b . Mitochondrial network analysis of MitoTracker Red live probed cells using the FIJI/ImageJ Mitochondria Analyzer plugin. Distribution of the mean mitochondrial area and mean mitochondrial perimeter in treated cells. Mitochondrial aspect ratio (length/width) was calculated using the plugin. Results are represented as mean ± SEM from ≥ 75 cells ( n = 3). The p -values are denoted numerically , ns = not significant, denoted by line in the graph, or from Untreated, by One-way ANOVA, Tukey honesty significant difference post-hoc testing. c . Quantitative analysis of mitochondrial membrane potential (ΔΨm) using tetramethylrhodamine ethyl ester (TMRE). Results of relative fluorescence are represented as mean ± SEM ( n = 3). ns = not significant from Untreated, by One-way ANOVA, Tukey honesty significant difference post-hoc testing. d . Effect of lysosomal protease inhibitor co-treatment with GBCAs on the metabolic capacity of exposed cells as a measure of cell viability. Data are represented as mean ± SEM ( n = 3 per group). The p -values are denoted numerically, ns = not significant from Untreated, by One-way ANOVA, Tukey honesty significant difference post-hoc testing. Protease inhibitor cocktail (PI) (P1860, Sigma-Aldrich). CA-074 methyl ester (CA-074Me) (S7420, SelleckChem)

Article Snippet: Mouse renal proximal tubular epithelial cells (MRPTEpiCs) (ScienCell Research Laboratories, #M4100-57) were cultured according to recommended protocols in complete epithelial cell medium-animal (c/EpiCM-a) (ScienCell Research Laboratories, #4131) and maintained at 37oC and 5% CO 2 .

Techniques: Inhibition, Labeling, Produced, Membrane, Fluorescence, Protease Inhibitor