normal rat kidney fibroblasts  (ATCC)

Journal: Cell Death Discovery

Article Title: Inhibition of epigenetic regulator UHRF1 attenuates renal fibrosis and retains transcription factor Krüppel-like factor 15 expression

doi: 10.1038/s41420-025-02549-y

Figure Lengend Snippet: A Masson staining (×200) and immunochemistry staining (×200) for α-SMA and Fibronectin on days 7 in UUO mice and on days 14 in UIR mice. Scale bar: 100 μm. B The bar graph summarizes the relative fibrotic area or α-SMA-positive and Fibronectin positive areas in each group. C , D Relative renal KLF15 protein expression in UUO mice or in UIR mice, analyzed by western blots. E , F Immunofluorescence double-labeling with antibodies to UHRF1 (Red) and PDFGRα + β (green) or AQP1 (green) (×200). The white arrow represents UHRF1 + fibroblasts (top panel) and UHRF1 + proximal tubular epithelial cells (bottom panel). Scale bar: 50 μm. n = 6 mice per group. Data were shown as mean ± SEM. Data were analyzed using unpaired Student’s t test. * P < 0.05, ** P < 0.01.

Article Snippet: Rat normal renal fibroblast (NRK-49F cells, ATCC) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin.

Techniques: Staining, Expressing, Western Blot, Immunofluorescence, Labeling

Journal: Cell Death Discovery

Article Title: Inhibition of epigenetic regulator UHRF1 attenuates renal fibrosis and retains transcription factor Krüppel-like factor 15 expression

doi: 10.1038/s41420-025-02549-y

Figure Lengend Snippet: Col1a2-Cre + /UHRF1 flox/flox mice (fibroblast-specific depletion of UHRF1) and their littermate controls received Sham or UUO operation for 7 days. A Generation of mice with fibroblast-specific depletion of UHRF1 using Cre-LoxP recombination system. B Genotyping of conditional knockout (cKO) mice (Col1a2-Cre + /UHRF1 flox/flox mice). C Illustration of experimental design to induce fibroblast-specific depletion of UHRF1 in UUO mice. D Relative renal UHRF1 protein expression analyzed by Western blots. E Immunofluorescence double-labeling with antibodies to UHRF1 (Red) and PDFGRα + β (green) (×200). The white arrow represents UHRF1 + fibroblasts. Scale bar: 50 μm. Masson staining ( F , H ) and immunochemistry staining for α-SMA and Fibronectin ( G , I ) in kidney sections (×200). Scale bar: 100 μm. ( J , K ) Western blots show the renal expression of α-SMA and Fibronectin. n = 6 mice per group. Data were shown as mean ± SEM. Data were analyzed using One-way ANOVA followed by Tukey. * P < 0.05, ** P < 0.01.

Article Snippet: Rat normal renal fibroblast (NRK-49F cells, ATCC) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin.

Techniques: Knock-Out, Expressing, Western Blot, Immunofluorescence, Labeling, Staining

Journal: Cell Death Discovery

Article Title: Inhibition of epigenetic regulator UHRF1 attenuates renal fibrosis and retains transcription factor Krüppel-like factor 15 expression

doi: 10.1038/s41420-025-02549-y

Figure Lengend Snippet: Col1a2-Cre + /UHRF1 flox/flox mice (fibroblast-specific depletion of UHRF1) and their littermate controls received Sham or UIR operation for 14 days. A Illustration of experimental design to induce fibroblast-specific depletion of UHRF1 in UIR mice. B Immunofluorescence double-labeling with antibodies to UHRF1 (Red) and PDFGRα + β (green) (×200). The white arrow represents UHRF1 + fibroblasts. Scale bar: 50 μm. C , D Masson staining and immunochemistry staining for α-SMA and Fibronectin in kidney sections (×200). Scale bar: 100 μm. n = 6 mice per group. Data were shown as mean ± SEM. Data were analyzed using One-way ANOVA followed by Tukey. * P < 0.05, ** P < 0.01.

Article Snippet: Rat normal renal fibroblast (NRK-49F cells, ATCC) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin.

Techniques: Immunofluorescence, Labeling, Staining

Journal: Cell Death Discovery

Article Title: Inhibition of epigenetic regulator UHRF1 attenuates renal fibrosis and retains transcription factor Krüppel-like factor 15 expression

doi: 10.1038/s41420-025-02549-y

Figure Lengend Snippet: A MeDIP-qPCR analysis shows the level of KLF15 promotor methylation. NRK-49F cells were transfected with UHRF1 siRNA or control siRNA before treatment of 10 ng/ml TGF-β1 for 48 h. n = 3 samples per group. B , C Col1a2-Cre + /UHRF1 flox/flox mice (fibroblast-specific depletion of UHRF1) and their littermate controls received Sham or UUO operation for 7 days. B Renal KLF15 promotor methylation levels were determined by MeDIP-qPCR. n = 4 mice per group. C Renal KLF15 protein levels were determined by Western blots. n = 6 mice per group. Data were shown as mean ± SEM. Data were analyzed using unpaired Student’s t test ( B ) or One-way ANOVA followed by Tukey ( A , C ). * P < 0.05, ** P < 0.01.

Article Snippet: Rat normal renal fibroblast (NRK-49F cells, ATCC) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin.

Techniques: Methylated DNA Immunoprecipitation, Methylation, Transfection, Control, Western Blot

Journal: Cell Death Discovery

Article Title: Inhibition of epigenetic regulator UHRF1 attenuates renal fibrosis and retains transcription factor Krüppel-like factor 15 expression

doi: 10.1038/s41420-025-02549-y

Figure Lengend Snippet: A Western blots showed the DNMT1 expression in NRK-49F cells transfected with UHRF1 siRNA or control siRNA with or without TGF-β1 treatment. B DNMT1 enzyme activity of NRK-49F cells transfected with UHRF1 siRNA or control siRNA with or without TGF-β1 treatment. C Co-IP analysis showed anti-UHRF1 antibody pulled down endogenous DNMT1. D PLA assay showed the interaction between UHRF1 and DNMT1 in NRK-49F cells with or without TGF-β1 treatment, presented by red fluorenscent signals (×400). Scale bar: 20 μm. E MeDIP-qPCR analysis showed inhibition of the interaction between UHRF1 and DNMT1 with NSC232003 prevented TGF-β1-induced KLF15 hypermethylation in NRK-49F cells. F Western blots demonstrated blocking effect of NSC232003 on TGF-β1-induced fibroblasts activation in NRK-49F cells. G NRK-49F cells infected with CRISPR-Cas9 lentivirus targeting KLF15 or scramble lentivirus were treated with TGF-β1 and NSC232003. Fibroblasts activation was assessed by immunoblots of α-SMA and Fibronectin. n = 3–6 samples per group. Data were shown as mean ± SEM. Data were analyzed using unpaired Student’s t test ( D , G ) or One-way ANOVA followed by Tukey ( A , B , E , F ). * P < 0.05, ** P < 0.01.

Article Snippet: Rat normal renal fibroblast (NRK-49F cells, ATCC) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin.

Techniques: Western Blot, Expressing, Transfection, Control, Activity Assay, Co-Immunoprecipitation Assay, Methylated DNA Immunoprecipitation, Inhibition, Blocking Assay, Activation Assay, Infection, CRISPR