Journal: Inflammation

Article Title: ZBP1 Mediates Renal Tubular Injury in Diabetic Nephropathy Through RIPK3-mediated Necroptosis

doi: 10.1007/s10753-025-02362-w

Figure Lengend Snippet: ZBP1 knockdown reduces HG-induced MTECs damage and necroptosi s in vitro. (a and b) The knockdown effect of ZBP1 was detected by real-time PCR and Western blot. (c) Western blot analysis of KIM1 in MTECs. (d) IF of KIM1 in MTECs. ZBP1 knockdown reduced KIM-1 expression in HG-induced MTECs. Scale bars = 50 μm. (e) Quantitative data and Western blotting of RIPK3, p-RIPK3, MLKL and p-MLKL. (f) Immunofluorescence analysis of p-MLKL in MTECs. Scale bars = 50 μm. (g) TEM revealed that HG increased MTEC volume, induced cell membrane rupture (blue puncta), cytoplasmic translucency (yellow puncta), organelle swelling (red puncta) and cytoplasmic content leakage. These pathological changes were attenuated in the ZBP1 knockdown group. Abbreviations: EV, empty vector; KD, knockdown; IHC, immunohistochemistry; HG, high glucose; IF, immunofluorescence; TEM, transmission electron microscopy. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse tubular epithelial cells (MTECs) (Procell, CP-M062) were cultured in low-glucose DMEM (Gibco, 11885092) supplemented with 8% fetal bovine serum (FBS; Gibco, 10270-106).

Techniques: Knockdown, In Vitro, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Immunofluorescence, Membrane, Plasmid Preparation, Immunohistochemistry, Transmission Assay, Electron Microscopy

Journal: Inflammation

Article Title: ZBP1 Mediates Renal Tubular Injury in Diabetic Nephropathy Through RIPK3-mediated Necroptosis

doi: 10.1007/s10753-025-02362-w

Figure Lengend Snippet: ZBP1 knockdown attenuates HG-induced inflammatory response and fibrosis in MTECs. (a) Real-time PCR analysis mRNA levels of IL-1β and MCP-1. (b) Quantitative data and Western blot analysis of p-p65 in MTECs stimulated with HG. (c) Immunofluorescence analysis of TNF-α in MTECs stimulated with HG. Scale bars = 25 μm. (d) Western blot showed that inhibition of ZBP1 expression could inhibit the expression of α-SMA and COL-I. (e) Immunofluorescence also showed the expression of α-SMA. Scale bars = 25 μm. Abbreviations: HG, high glucose; EV, empty vector; KD, knockdown. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse tubular epithelial cells (MTECs) (Procell, CP-M062) were cultured in low-glucose DMEM (Gibco, 11885092) supplemented with 8% fetal bovine serum (FBS; Gibco, 10270-106).

Techniques: Knockdown, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence, Inhibition, Expressing, Plasmid Preparation

Journal: Inflammation

Article Title: ZBP1 Mediates Renal Tubular Injury in Diabetic Nephropathy Through RIPK3-mediated Necroptosis

doi: 10.1007/s10753-025-02362-w

Figure Lengend Snippet: ZBP1 knockdown attenuates AGEs-induced MTECs damage and necroptosis in vitro. (a and b) Western blot and real-time PCR of KIM-1 in MTECs. (c) Quantitative data and Western blotting of RIPK3, p-RIPK3, MLKL and p-MLKL. (d) Immunofluorescence analysis of p-MLKL in MTECs. Scale bars = 25 μm. (e) Real-time PCR analysis mRNA levels of IL-1β and TNF-α. (f) Quantitative data and Western blot analysis of p-p65 in MTECs stimulated with AGEs. (g) Western blot showed that inhibition of ZBP1 expression could inhibit the expression of α-SMA and COL-I. Abbreviations: AGEs, advanced glycation end-products; EV, empty vector; KD, knockdown. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse tubular epithelial cells (MTECs) (Procell, CP-M062) were cultured in low-glucose DMEM (Gibco, 11885092) supplemented with 8% fetal bovine serum (FBS; Gibco, 10270-106).

Techniques: Knockdown, In Vitro, Western Blot, Real-time Polymerase Chain Reaction, Immunofluorescence, Inhibition, Expressing, Plasmid Preparation

Journal: Inflammation

Article Title: ZBP1 Mediates Renal Tubular Injury in Diabetic Nephropathy Through RIPK3-mediated Necroptosis

doi: 10.1007/s10753-025-02362-w

Figure Lengend Snippet: ZBP1 interacts with RIPK3 in HG-induced MTECs. ( a ) Western blot showed the protein levels of RIPK3, p-RIPK3, MLKL and p-MLKL. ( b ) Immunofluorescence images showed that ZBP1 knockdown and GSK’872 reduced the phosphorylation and membrane translocation of MLKL in HG-induced MTECs. ( c ) Western blot revealed that overexpressing RIPK3 negated the protective effects of ZBP1 knockdown on necroptosis. ( d - f ) Co-IP, SPR and molecular docking of ZBP1 and RIPK3. ( e ) Molecular docking of ZBP1 and RIPK3. Docking scale = 6.88 Kcal/mol. Abbreviations: EV, empty vector; KD, knockdown; HG, high glucose; IF, immunofluorescence; IP, immunoprecipitation; SPR, surface plasmon resonance. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Mouse tubular epithelial cells (MTECs) (Procell, CP-M062) were cultured in low-glucose DMEM (Gibco, 11885092) supplemented with 8% fetal bovine serum (FBS; Gibco, 10270-106).

Techniques: Western Blot, Immunofluorescence, Knockdown, Phospho-proteomics, Membrane, Translocation Assay, Co-Immunoprecipitation Assay, Plasmid Preparation, Immunoprecipitation, SPR Assay

Journal: Inflammation

Article Title: ZBP1 Mediates Renal Tubular Injury in Diabetic Nephropathy Through RIPK3-mediated Necroptosis

doi: 10.1007/s10753-025-02362-w

Figure Lengend Snippet: ETS1 targets and promotes the transcription of ZBP1. (a) The Venn diagram showed the transcription factors that the database predicts regulate ZBP1 expression. (b) Pearson Correlation Analysis of transcription factors and ZBP1. (c) Real-time PCR for transcription factors in db/db mice. (d) Western blot results showed that ETS1 was knocked down by siRNA. (e) Western blot results revealed that knocking down ETS1 decreased ZBP1 expression in HG-stimulated MTECs. (f and g) ETS1 binds to the ZBP1 promoter, and primer 3 binds most strongly relative to other sites. Abbreviations: HG, high glucose; EV, empty vector; KD, knockdown. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse tubular epithelial cells (MTECs) (Procell, CP-M062) were cultured in low-glucose DMEM (Gibco, 11885092) supplemented with 8% fetal bovine serum (FBS; Gibco, 10270-106).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Plasmid Preparation, Knockdown