rat kidney proximal tubular epithelial nrk52e cells  (ATCC)

Journal: iScience

Article Title: Injured tubular epithelia-derived CCN1 promotes the mobilization of fibroblasts toward injury sites after kidney injury

doi: 10.1016/j.isci.2025.112176

Figure Lengend Snippet: PT-specific transcriptomics reveal Ccn1 up-regulation in the early phase after cisplatin-induced kidney injury in vivo (A) Experimental scheme. Bigenic mice (SLC34a1GCE × R26tdTomato) received intraperitoneal injections of cisplatin (15 mg/kg) and tamoxifen (3 mg/kg at each time point) as indicated. (B) Changes in BUN after the administration of cisplatin. n = 4–5 per group. (C) Isolation of tdTomato+ tubular epithelial cells using FACS. (D) Histological analysis of the kidney after the cisplatin injection. PAS staining of kidney sections and immunostaining of LTL and KIM1. The scale bars indicate 50 μm in PAS staining, 100 μm in low-power field pictures and 20 μm in high-power field pictures in immunostaining. (E) Time-course analyses of qPCR of RNA from the whole kidney for the representative markers of tubular injury ( Havcr1 ), profibrotic factors ( Ccn1 , Tgfb1 , Ctgf , and Pdgfb ), myofibroblast ( Acta2 ), mesenchymal cell ( Vim ), and extracellular matrix ( Col1a1 and Fn1 ). n = 4–5 per group. (F) Time-course analyses of qPCR of RNA from the isolated tdTomato+ tubular epithelial cells. n = 4–5 per group. For all groups, data are means ± SEM. Statistical analyses were performed by unpaired t test for comparisons to day 0 (B, E, F). ∗ p < 0.05 vs. day 0.

Article Snippet: Rat kidney epithelial cells (NRK52E cells) , The JCRB Cell Bank , N/A.

Techniques: In Vivo, Isolation, Injection, Staining, Immunostaining

Journal: iScience

Article Title: Injured tubular epithelia-derived CCN1 promotes the mobilization of fibroblasts toward injury sites after kidney injury

doi: 10.1016/j.isci.2025.112176

Figure Lengend Snippet: The PT-specific knockout of CCN1 inhibits the accumulation of fibroblasts at injured tubules in cisplatin nephrotoxicity in vivo (A) Experimental scheme. Tubular-specific CCN1 knockout mice (CCN1Flox/Flox SLC34a1GCE x R26tdTomato: CCN-KO) received intraperitoneal injections of cisplatin (15 mg/kg) and tamoxifen (3 mg/kg at each time point) as indicated. (B) Histological analysis of the kidney after the cisplatin injection by immunostaining of KIM1. (C) RT-PCR of the Ccn1 and Havcr1 genes in sorted tdTomato+ tubular epithelial cells. (D) Changes in BUN after the administration of cisplatin. n = 7–8 per group. (E) Representative co-immunofluorescence images of KIM1 and pFAK, PDGFRβ and pFAK, and PDGFRβ and γH2AX at 4 days after cisplatin injection. (F) Representative images of Sirius red staining at 14 days after cisplatin injection. (G) Semi-quantitative fibrosis scores. n = 9 WT Cis (−), n = 8 WT Cis (+), n = 7 CCN1-KO Cis (+). (H) qPCR of RNA from whole kidneys as representative markers of myofibroblasts ( Acta2 ), fibroblasts ( Pdgfrb ), profibrotic factor ( Tgfb1 ), macrophage ( Cd68 ), tubular injury ( Havcr1 ), tubular integrity ( Lrp2 ), and extracellular matrix ( Col1a1 and Fn1 ). n = 5 WT Cis (−), n = 8 WT Cis (+), n = 7 CCN1-KO Cis (+). The scale bars indicate 100 μm in (B), 50 μm in low-power field pictures, and 20 μm in high-power field pictures in (E). The scale bars indicate 50 μm in (F). For all groups, data are means ± SEM. Statistical analyses were performed by unpaired t test for comparisons of two variables and by ANOVA and Dunnett’s post hoc test for comparisons of multiple variables. ∗ p < 0.05.

Article Snippet: Rat kidney epithelial cells (NRK52E cells) , The JCRB Cell Bank , N/A.

Techniques: Knock-Out, In Vivo, Injection, Immunostaining, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Staining

Journal: iScience

Article Title: Injured tubular epithelia-derived CCN1 promotes the mobilization of fibroblasts toward injury sites after kidney injury

doi: 10.1016/j.isci.2025.112176

Figure Lengend Snippet: The PT-specific knockout of CCN1 inhibits the accumulation of fibroblasts at injured tubules and subsequent tissue fibrosis in IRI in vivo (A) Experimental scheme. Tubular-specific CCN1 knockout mice (CCN1Flox/Flox SLC34a1GCE x R26tdTomato: CCN-KO) received IRI (35 min) and tamoxifen (3 mg/kg at each time point) as indicated. (B) Histological analysis of the kidney at 1 day after IRI by immunostaining of KIM1. (C) RT-PCR of the Ccn1 and Havcr1 genes in sorted tdTomato+ tubular epithelial cells. (D) Representative co-immunofluorescence images of KIM1 and pFAK, and PDGFRβ and pFAK at 1 day after IRI. (E) Representative images of Sirius red staining. (F) Semi-quantitative fibrosis scores at 14 days after IRI. n = 9 WT IRI (−), n = 8 WT IRI (+), n = 9 CCN1-KO IRI (−), n = 8 CCN1-KO IRI (+). (G) qPCR of RNA from whole kidneys as representative markers of tubular injury ( Havcr1 ), tubular integrity ( Lrp2 ), chemokine ( Ccl2 ), macrophage ( Cd68 ), fibroblasts ( Pdgfrb ), myofibroblasts ( Acta2 ), extracellular matrix ( Col1a1 and Fn1 ), and profibrotic factor ( Tgfb1 ). n = 9 WT IRI (−), n = 8 WT IRI (+), n = 8 CCN1-KO IRI (−), n = 7 CCN1-KO IRI (+). The scale bars indicate 100 μm in (B), 50 μm in low-power field pictures, and 20 μm in high-power field pictures in (D). The scale bars indicate 50 μm in (E). For all groups, data are means ± SEM. Statistical analyses were performed by paired t test for comparisons of IRI kidney and contralateral kidney (CLK) and by ANOVA and Dunnett’s post hoc test for comparisons of multiple variables. ∗ p < 0.05.

Article Snippet: Rat kidney epithelial cells (NRK52E cells) , The JCRB Cell Bank , N/A.

Techniques: Knock-Out, In Vivo, Immunostaining, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Staining

Journal: iScience

Article Title: Injured tubular epithelia-derived CCN1 promotes the mobilization of fibroblasts toward injury sites after kidney injury

doi: 10.1016/j.isci.2025.112176

Figure Lengend Snippet:

Article Snippet: Rat kidney epithelial cells (NRK52E cells) , The JCRB Cell Bank , N/A.

Techniques: Plasmid Preparation, Virus, CRISPR, Recombinant, Inhibition, Negative Control, Saline, Western Blot, Proliferation Assay, Sample Prep, Software, Microscopy

Journal: International Journal of Molecular Sciences

Article Title: PPAR Pan Agonist MHY2013 Alleviates Renal Fibrosis in a Mouse Model by Reducing Fibroblast Activation and Epithelial Inflammation

doi: 10.3390/ijms24054882

Figure Lengend Snippet: MHY2013 suppresses LPS-induced inflammatory responses in NRK52E cells. ( A ) Relative mRNA levels of inflammation-related genes ( Ccl2 , Ccl5 , and Cxcl1 ) in LPS-treated NRK52E cells with or without MHY2013. # p < 0.05 vs. control group. * p < 0.05 vs. LPS group. ( B ) NF-κB transcriptional activity was measured under LPS-treated condition with or without MHY2013 treatment. *** p < 0.001 vs. non-treated group. ### p < 0.001 vs. LPS-treated group. ( C ) Relative mRNA levels of inflammation-related genes ( Ccl2, Ccl5 , and Cxcl1 ) in LPS-treated NRK52E cells with or without PPARβ expression.

Article Snippet: NRK52E rat-kidney epithelial cells were purchased from ATCC (CRL-1571) and grown in DMEM supplemented with 10% FBS and 1% penicillin.

Techniques: Control, Activity Assay, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Nifedipine Modulates Renal Lipogenesis via the AMPK-SREBP Transcriptional Pathway

doi: 10.3390/ijms20071570

Figure Lengend Snippet: The effect of nifedipine on the cell viability, and intracellular lipid accumulation in NRK52E tubular cells. ( a ) The nifedipine treated cells (15, 30 μM) showed decreased viability compared to control ( p < 0.01) in 24 or 48 h of MTT assays. ( n = 3) ( b ) The nifedipine treated cells (30 μM) and oleic acid (150 μM) showed an increasing effect on quantification of Oil Red O staining comparing to control ( p < 0.01) in 24 or 48 h. ( n = 3) ( c ) The microscopic Oil Red O staining of NRK52E (magnification, 400×) treated with nifedipine and oleic acid are also shown: ( top ) 24 h; and ( bottom ) 48 h. p -values ≤ 0.05 (marked as **) were considered statistically significant. The bar size in ( c ) is 100 μM.

Article Snippet: The normal rat kidney epithelial-derived cell line NRK52E (CRL-1571) was obtained from the Bioresource Collection and Research Center, Food Industry Research Development Institute, Hsinchu, Taiwan.

Techniques: Staining