normal rat kidney fibroblasts  (ATCC)

Journal: Renal Failure

Article Title: Ubiquitin-specific protease 11 facilitates the activation and proliferation of renal interstitial fibroblasts through epidermal growth factor receptor signaling pathways

doi: 10.1080/0886022X.2026.2666452

Figure Lengend Snippet: USP11 is highly upregulated in UA-stimulated RIF. In vitro , we stimulated NRK49F with UA in dose-dependent manner (0, 200, 400, 800 μM) and in time-dependent manner (0, 12, 24, 36 hours). Western blot analysis of USP11, α-SMA and Collagen I protein expressions in each group were conducted, with normalization to GAPDH (A, C). Quantitative analysis of USP11, α-SMA and Collagen I protein levels in each group (B, D). Data were expressed as mean± SD ( n = 4 for each group). N.S.: no significant difference. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Rat renal interstitial fibroblast (NRK49F) cells were obtained from ATCC (Manassas, VA).

Techniques: In Vitro, Western Blot

Journal: Renal Failure

Article Title: Ubiquitin-specific protease 11 facilitates the activation and proliferation of renal interstitial fibroblasts through epidermal growth factor receptor signaling pathways

doi: 10.1080/0886022X.2026.2666452

Figure Lengend Snippet: USP11 contributes to RIF activation in the UA-stimulated NRK49F. NRK49F cells were seeded to 70–80% confluence in the antibiotic-free medium and grown followed by transfection with USP11 siRNA or USP11-pcDNA 3.0 plasmid. After transfection, the medium was changed to DMEM with F12 containing 0.5% FBS for starvation and then cells were incubated with or without uric acid (800 μM) for an additional 36 hours before being harvested for analysis. Western blot analysis of USP11, α-SMA and Collagen I protein expressions in each group were conducted, with normalization to GAPDH (A, F, H). Quantitative analysis of USP11, α-SMA and Collagen I protein levels in each group (B, G, I). Representative images and quantitative analysis of immunofluorescence staining for Fibronectin and Vimentin with DAPI nuclear counterstaining in each group (C). Then NRK49F cells were starved for 24 h with DMEM containing 0.5% FBS before they were exposed to uric acid in the presence or absence of MTX (1, 5, and 10 μM). Western blot analysis of USP11, α-SMA and Collagen I protein expressions in each group were conducted, with normalization to GAPDH (D). Quantitative analysis of USP11, α-SMA and Collagen I protein levels in each group (E). Data were expressed as mean± SD ( n = 4 for each group). N.S.: no significant difference. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Scale bars = 100μm.

Article Snippet: Rat renal interstitial fibroblast (NRK49F) cells were obtained from ATCC (Manassas, VA).

Techniques: Activation Assay, Transfection, Plasmid Preparation, Incubation, Western Blot, Immunofluorescence, Staining

Journal: Renal Failure

Article Title: Ubiquitin-specific protease 11 facilitates the activation and proliferation of renal interstitial fibroblasts through epidermal growth factor receptor signaling pathways

doi: 10.1080/0886022X.2026.2666452

Figure Lengend Snippet: USP11 directly interacts with EGFR and stabilizes its protein level. Immunofluorescence co-staining of USP11 and EGFR in the UA-stimulated NRK49F (A). Co-IP assay in NRK49F cells. Flag-tagged USP11 and Myc-tagged EGFR were immunoprecipitated using an anti-Flag/Myc antibody, and the presence of Myc-tagged EGFR and Flag-tagged USP11 in the precipitates was detected by immunoblotting (B, C). Western blot analysis of USP11 and EGFR protein expressions in each group were conducted, with normalization to GAPDH (D). Quantitative analysis of EGFR protein levels in each group (E). Data were expressed as mean± SD ( n = 4 for each group). N.S.: no significant difference, *** p < 0.001. Scale bars = 50μm.

Article Snippet: Rat renal interstitial fibroblast (NRK49F) cells were obtained from ATCC (Manassas, VA).

Techniques: Immunofluorescence, Staining, Co-Immunoprecipitation Assay, Immunoprecipitation, Western Blot

Journal: Renal Failure

Article Title: Ubiquitin-specific protease 11 facilitates the activation and proliferation of renal interstitial fibroblasts through epidermal growth factor receptor signaling pathways

doi: 10.1080/0886022X.2026.2666452

Figure Lengend Snippet: USP11 activates EGFR signaling pathway in the UA-stimulated NRK49F. Western blot analysis of p-EGFR and EGFR protein expressions in each group were conducted, with normalization to GAPDH (A, C, E). Quantitative analysis of p-EGFR and EGFR protein levels in each group (B, D, F). Data were expressed as mean ± SD (n = 4 for each group). N.S.: no significant difference. *** p < 0.001, **** p < 0.0001.

Article Snippet: Rat renal interstitial fibroblast (NRK49F) cells were obtained from ATCC (Manassas, VA).

Techniques: Western Blot

Journal: Renal Failure

Article Title: Ubiquitin-specific protease 11 facilitates the activation and proliferation of renal interstitial fibroblasts through epidermal growth factor receptor signaling pathways

doi: 10.1080/0886022X.2026.2666452

Figure Lengend Snippet: USP11 is involved in the activation of EGFR signaling pathway in the UA-stimulated NRK49F. To fully demonstrate the relationship between USP11 and EGFR signaling, we stimulated starved NRK49F cells with EGF (5 ng/ml) in the presence or absence of USP11 siRNA or treated with gefitinib (1 nM and 5 nM), a highly selective EGFR inhibitor in the presence of USP11 pcDNA 3.0 plasmid for an additional 36 hours. Western blot analysis of p-EGFR and EGFR protein expressions in each group were conducted, with normalization to GAPDH (A, E). Quantitative analysis of p-EGFR and EGFR protein levels in each group (B, F). Western blot analysis of USP11, α-SMA and Collagen I protein expressions in each group were conducted, with normalization to GAPDH (C, G). Quantitative analysis of USP11, α-SMA and Collagen I protein levels in each group (D, H). Data were expressed as mean ± SD ( n = 4 for each group). N.S.: no significant difference. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Rat renal interstitial fibroblast (NRK49F) cells were obtained from ATCC (Manassas, VA).

Techniques: Activation Assay, Plasmid Preparation, Western Blot

Journal: Renal Failure

Article Title: Ubiquitin-specific protease 11 facilitates the activation and proliferation of renal interstitial fibroblasts through epidermal growth factor receptor signaling pathways

doi: 10.1080/0886022X.2026.2666452

Figure Lengend Snippet: Inhibition of USP11 attenuates the proliferation and migration in the UA-stimulated NRK49F. Photomicrographs of migrating cells in wound healing assay were taken at 0h and 36h (A). The migratory rate was calculated as (A-B)/A*100%, where A and B reflect the width of the wound at 0h and 36h respectively (B). The CCK-8 proliferation kit was used according to the manufacturer’s instructions and the final optical density values were read at 450 nm (C). Representative images and quantitative analysis of immunofluorescence staining for Ki67 with DAPI nuclear counterstaining in each group (D, E). Western blot analysis of PCNA and Cyclin E protein expressions in each group were conducted, with normalization to GAPDH (F, H). Quantitative analysis of PCNA and Cyclin E protein levels in each group (G, I). Data were expressed as mean± SD ( n = 4 for each group). N.S., no significant difference. * p < 0.05, *** p < 0.001, **** p < 0.0001. Scale bars = 500μm (A) and 50μm (D).

Article Snippet: Rat renal interstitial fibroblast (NRK49F) cells were obtained from ATCC (Manassas, VA).

Techniques: Inhibition, Migration, Wound Healing Assay, CCK-8 Assay, Immunofluorescence, Staining, Western Blot

Journal: Frontiers in Medicine

Article Title: Identification of novel protein biomarkers for knee osteoarthritis by integrating human plasma proteome: evidence from Mendelian randomization and preliminary in vitro investigation

doi: 10.3389/fmed.2026.1745114

Figure Lengend Snippet: (A) Synovial fibroblast vimentin immunofluorescence detection results; (B) macrophage CD68 flow cytometry detection results.

Article Snippet: Rat peritoneal macrophages (MØs) (Cat. no. CP-R158) and rat synovial fibroblasts (SF) (Cat. no. CP-R329) were supplied by Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Immunofluorescence, Flow Cytometry

Journal: Frontiers in Medicine

Article Title: Identification of novel protein biomarkers for knee osteoarthritis by integrating human plasma proteome: evidence from Mendelian randomization and preliminary in vitro investigation

doi: 10.3389/fmed.2026.1745114

Figure Lengend Snippet: Galectin-3 inducing secretion of pro-inflammatory cytokines in macrophages and synovial fibroblasts.

Article Snippet: Rat peritoneal macrophages (MØs) (Cat. no. CP-R158) and rat synovial fibroblasts (SF) (Cat. no. CP-R329) were supplied by Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques:

Journal: Frontiers in Medicine

Article Title: Identification of novel protein biomarkers for knee osteoarthritis by integrating human plasma proteome: evidence from Mendelian randomization and preliminary in vitro investigation

doi: 10.3389/fmed.2026.1745114

Figure Lengend Snippet: (A) Expression of PRTN3/MPO/P-NF-κB p65 in macrophage; (B) expression of PRTN3/MPO/P-NF-κB p65 in synovial fibroblasts.

Article Snippet: Rat peritoneal macrophages (MØs) (Cat. no. CP-R158) and rat synovial fibroblasts (SF) (Cat. no. CP-R329) were supplied by Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing