rat renal tubular epithelial cell line  (ATCC)

Journal: Acta Pharmaceutica Sinica. B

Article Title: Targeting renal tubular epithelial cells via a dual functionalized oligosaccharide self-assembly in the management of acute and chronic kidney diseases

doi: 10.1016/j.apsb.2025.09.017

Figure Lengend Snippet: Schematic illustration of an active and passive dual-targeted delivery system (COA-SA/CLT-siSnail) for the codelivery of small molecule drugs and therapeutic nucleic acids to RTECs in the treatment of renal disease.

Article Snippet: Rat renal tubular epithelial cells NRK-52E, rat fibroblasts NRK-49F, and human proximal tubular epithelial cells HK-2 were purchased from American Type Culture Collection (Manassas, VA, USA), mouse renal tubular epithelial cells TCMK-1 were obtained from Dr. Li Yanping’s laboratory, and maintained in Dulbecco’s modified Eagle’s medium (DMEM) with high glucose supplemented with 10% ( v / v ) fetal bovine serum (FBS), streptomycin (100 μg/mL) and penicillin (100 U/mL) (Thermo Fisher, USA).

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Targeting renal tubular epithelial cells via a dual functionalized oligosaccharide self-assembly in the management of acute and chronic kidney diseases

doi: 10.1016/j.apsb.2025.09.017

Figure Lengend Snippet: Cellular uptake and internalization pathway of COA-SA/Cy5-siSnail micelles. (A) LSCM analysis cellular uptake of free Cy5-siSnail, COA-SA/Cy5-siSnail, Lipo6000/Cy5-siSnail in NRK-52E and HK-2 cells. Scale bars, 20 μm. (B) Flow cytometry analysis shows a positive rate of different preparations in RTECs. (C) Internalization of COA-SA/siSnail micelles in NRK-52E and HK-2 cells pretreated with 4 °C, methyl- β -cyclodextrin (M- β -CD), nystatin (Nys), 5-( N , N -dimethyl)-amiloride hydrochloride (DMA) and chlorpromazine (Chlo), respectively. ∗∗∗∗ P < 0.0001 vs . control. (D) Internalization of COA-SA/siSnail micelles in NRK-52E and HK-2 cells pretreated with EDTA, Gentamicin, Megalin antibody, and chitosan oligosaccharide (COS), respectively. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001 vs . control. Data represent mean ± SD ( n = 3). (E) LSCM shows the endosome/lysosome escape of COA-SA/Cy5-siSnail micelles in NRK-52E and HK-2 cells. Scale bars, 10 μm. (F) Inhibitory effects of COA-SA/CLT-siSnail micelles on the viability of RTECs. Data represent mean ± SD ( n = 6).

Article Snippet: Rat renal tubular epithelial cells NRK-52E, rat fibroblasts NRK-49F, and human proximal tubular epithelial cells HK-2 were purchased from American Type Culture Collection (Manassas, VA, USA), mouse renal tubular epithelial cells TCMK-1 were obtained from Dr. Li Yanping’s laboratory, and maintained in Dulbecco’s modified Eagle’s medium (DMEM) with high glucose supplemented with 10% ( v / v ) fetal bovine serum (FBS), streptomycin (100 μg/mL) and penicillin (100 U/mL) (Thermo Fisher, USA).

Techniques: Flow Cytometry, Control

Journal: bioRxiv

Article Title: In-vitro Knockdown and Pharmacological Inhibition of RIPK2 Attenuates Calcium Oxalate–Nephrocalcinosis Associated Inflammation and Oxidative Stress in NRK-52E and Primary Renal Epithelial Cells

doi: 10.1101/2025.07.13.664563

Figure Lengend Snippet: (A–B) Representative western blot and quantitative analysis of RIPK2/RICK protein expression in kidneys of CaOx-induced CKD mice (n=4). (C) RIPK2 mRNA expression in kidneys of CaOx-induced CKD mice (n=5). (D–E) Representative western blot and quantitative analysis of RIPK2/RICK protein expression in kidneys of adenine-induced nephrolithiasis mice (n=2). (F) RIPK2 mRNA expression in kidneys of adenine-induced nephrolithiasis mice (n=5). (G-J) Effect of HG, H • O • , adenine, and COM on RIPK2/RICK mRNA expression in NRK-52E cells. (K and P) NRK-52E cells and primary RTECs were exposed to different concentration of COM for 24 h. then 100 µg/mL dose was selected for further studies. (L and Q) Representative western blots of RICK expression in NRK-52E cells and primary RTECs respectively. (M and R) Quantitative analysis of western blots RICK/GAPDH (n=5). (N-O and S-T) Immunofluorescence expression of RICK and relative fluorescence intensity (n=4) respective to NRK-52E cells and primary RTECs. Values are expressed as mean ± SD. Unpaired t-test was used for statistical analysis (A-J). *p < 0.05, **p < 0.01, ***p < 0.001 vs respective control groups. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs respective control groups.

Article Snippet: NRK-52E rat renal tubular epithelial cells, procured from the National Centre for Cell Science (NCCS, Pune, India), were maintained in DMEM with high glucose (4.5g/L), supplemented with 10% FBS and 1% penicillin-streptomycin antibiotic cocktail.

Techniques: Western Blot, Expressing, Concentration Assay, Immunofluorescence, Fluorescence, Control

Journal: bioRxiv

Article Title: In-vitro Knockdown and Pharmacological Inhibition of RIPK2 Attenuates Calcium Oxalate–Nephrocalcinosis Associated Inflammation and Oxidative Stress in NRK-52E and Primary Renal Epithelial Cells

doi: 10.1101/2025.07.13.664563

Figure Lengend Snippet: Gene silencing effect of RICK siRNA on RICK protein and mRNA levels in NRK-52E cells were detected 24 D h after transfection. (A-B) Representative western blots of RICK and Quantitative analysis (n=3). (C) RICK mRNA expression (n=6). (D-E) Immunofluorescence expression of RICK and relative fluorescence intensity in NRK-52E cells (n=4). (F-G) Immunofluorescence expression of RICK and relative fluorescence intensity in primary RTECs (n=4). (H) RICK mRNA expression in primary RTECs (n=6). Values are expressed as mean ± SD. * p < 0.05 and ** p < 0.01 vs respective control and siCtrl groups.

Article Snippet: NRK-52E rat renal tubular epithelial cells, procured from the National Centre for Cell Science (NCCS, Pune, India), were maintained in DMEM with high glucose (4.5g/L), supplemented with 10% FBS and 1% penicillin-streptomycin antibiotic cocktail.

Techniques: Transfection, Western Blot, Expressing, Immunofluorescence, Fluorescence, Control

Journal: bioRxiv

Article Title: In-vitro Knockdown and Pharmacological Inhibition of RIPK2 Attenuates Calcium Oxalate–Nephrocalcinosis Associated Inflammation and Oxidative Stress in NRK-52E and Primary Renal Epithelial Cells

doi: 10.1101/2025.07.13.664563

Figure Lengend Snippet: After 24 h of siRNA transfection, COM was exposed to NRK-52E cells for additional 24 h. (A-B) FACS-DCFDA assay and Fura-2AM assay for ROS production and Intracellular Ca 2+ level respectively in NRK-52E cells (n=5). (C-D) Percent (%) ROS production and % intracellular Ca 2+ level in NRK-52E cells. (E) CAT mRNA expression level in NRK-52E cells (n=6). (F-G) FACS-DCFDA assay and Fura-2AM assay for ROS production and Intracellular Ca 2+ level respectively in primary RTECs (n=5). (H-I) % ROS production and % intracellular Ca 2+ level in primary RTECs. (J) CAT mRNA expression level in primary RTECs (n=6). Values are expressed as mean ± SD. * p < 0.05 and *** p < 0.001 vs respective control and siCtrl groups; # p < 0.05, ## p < 0.01, and ### p < 0.001 vs respective COM groups.

Article Snippet: NRK-52E rat renal tubular epithelial cells, procured from the National Centre for Cell Science (NCCS, Pune, India), were maintained in DMEM with high glucose (4.5g/L), supplemented with 10% FBS and 1% penicillin-streptomycin antibiotic cocktail.

Techniques: Transfection, Expressing, Control

Journal: bioRxiv

Article Title: In-vitro Knockdown and Pharmacological Inhibition of RIPK2 Attenuates Calcium Oxalate–Nephrocalcinosis Associated Inflammation and Oxidative Stress in NRK-52E and Primary Renal Epithelial Cells

doi: 10.1101/2025.07.13.664563

Figure Lengend Snippet: (A) Immunofluorescence expression of NF-κB, TNF-α, IL-1β, IL-6, and IL-10 in primary RTECs. (B-F) Relative fluorescence intensity of NF-κB, TNF-α, IL-1β, IL-6, and IL-10 (n=4). (G-K) The mRNA expression levels of IL-10, IL-6, IL-1β, TNF-α, and NF-κB in primary RTECs respectively (n=6). Values are expressed as mean ± SD. * p < 0.05 and *** p < 0.001 vs respective control groups; # p < 0.05, ## p < 0.01, and ### p < 0.001 vs respective COM groups.

Article Snippet: NRK-52E rat renal tubular epithelial cells, procured from the National Centre for Cell Science (NCCS, Pune, India), were maintained in DMEM with high glucose (4.5g/L), supplemented with 10% FBS and 1% penicillin-streptomycin antibiotic cocktail.

Techniques: Immunofluorescence, Expressing, Fluorescence, Control

Journal: bioRxiv

Article Title: In-vitro Knockdown and Pharmacological Inhibition of RIPK2 Attenuates Calcium Oxalate–Nephrocalcinosis Associated Inflammation and Oxidative Stress in NRK-52E and Primary Renal Epithelial Cells

doi: 10.1101/2025.07.13.664563

Figure Lengend Snippet: (A) TMRE dye was used for mitochondrial membrane potential by FACS in primary RTECs. (B) TMRE (%) positive cells (n=5). (C) The primary RTECs were stained with Annexin • -FITC/PI and analyzed by flow cytometry to determine the apoptosis population (n=5). (D) Quantitative data for the percentage of Healthy (Q3) early (Q4), late (Q2), and necrotic cells. (E) Caspase-3 mRNA expression level (n=6). (F) Immunofluorescence expression of caspase-3. (G) Relative fluorescence intensity of caspase-3 in primary RTECs (n=4). Values are expressed as mean ± SD. ** p < 0.01 and *** p < 0.001 vs respective control groups; # p < 0.05 and ## p < 0.01 vs respective COM groups.

Article Snippet: NRK-52E rat renal tubular epithelial cells, procured from the National Centre for Cell Science (NCCS, Pune, India), were maintained in DMEM with high glucose (4.5g/L), supplemented with 10% FBS and 1% penicillin-streptomycin antibiotic cocktail.

Techniques: Membrane, Staining, Flow Cytometry, Expressing, Immunofluorescence, Fluorescence, Control

Journal: bioRxiv

Article Title: In-vitro Knockdown and Pharmacological Inhibition of RIPK2 Attenuates Calcium Oxalate–Nephrocalcinosis Associated Inflammation and Oxidative Stress in NRK-52E and Primary Renal Epithelial Cells

doi: 10.1101/2025.07.13.664563

Figure Lengend Snippet: (A and G) Sirius red staining for collagen in NRK-52E cells and primary RTECs respectively. (B and H) H and E staining in NRK-52E cells and primary RTECs respectively. Result clearly shows the decreased in cell integrity, morphology, and viability in NRK-52E cells and primary RTECs exposed COM and increased collagen secretion and deposition (Black arrow indicate the collagen and white arrow indicate the abnormal structure of cells), whereas in RICK Gene Silencing and inhibition groups reversed collagen secretion, abnormality, integrity, and nuclear condensation. (C and I) Representative bright field CaOx crystal adhesion images in NRK-52E and primary RTECs respectively (orange arrow indicate the CaOx crystals). (D and J) Collagen deposition (%) (n=4). (E and K) Relative mean area of cells (n=4). (F and L) Crystal deposition in % area (n=4). Magnification × D 200. Values are expressed as mean ± SD. * p < 0.05 and *** p < 0.001 vs respective control and siCtrl groups; # p < 0.05 and ## p < 0.01 vs respective COM groups.

Article Snippet: NRK-52E rat renal tubular epithelial cells, procured from the National Centre for Cell Science (NCCS, Pune, India), were maintained in DMEM with high glucose (4.5g/L), supplemented with 10% FBS and 1% penicillin-streptomycin antibiotic cocktail.

Techniques: Staining, Inhibition, Control

Journal: bioRxiv

Article Title: In-vitro Knockdown and Pharmacological Inhibition of RIPK2 Attenuates Calcium Oxalate–Nephrocalcinosis Associated Inflammation and Oxidative Stress in NRK-52E and Primary Renal Epithelial Cells

doi: 10.1101/2025.07.13.664563

Figure Lengend Snippet: (A) Immunofluorescence expression of TGF-β, E-cadherin, and Vimentin in NRK-52E cells. (B-D) Relative fluorescence intensity of TGF-β, E-cadherin, and Vimentin (n=4). (E-I) The mRNA expression levels of TGF-β, Vimentin, Collagen-IV, E-cadherin, and α-SMA in primary RTECs respectively (n=6). Values are expressed as mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs respective control groups; # p < 0.05 and ## p < 0.01 vs respective COM groups.

Article Snippet: NRK-52E rat renal tubular epithelial cells, procured from the National Centre for Cell Science (NCCS, Pune, India), were maintained in DMEM with high glucose (4.5g/L), supplemented with 10% FBS and 1% penicillin-streptomycin antibiotic cocktail.

Techniques: Immunofluorescence, Expressing, Fluorescence, Control