kidney injury molecule 1 kim1  (Elabscience Biotechnology)

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Proprotein convertase subtilisin/kexin type 9 contributes to cisplatin-induced acute kidney injury by interacting with cyclase-associated protein 1 to promote megalin lysosomal degradation.

doi: 10.1016/j.bbamcr.2025.119984

Figure Lengend Snippet: Fig. 1. PCSK9 is upregulated in mouse renal cortex in cisplatin-induced AKI. (A) Representative images of co-immunofluorescence staining of PCSK9 with AQP1 or Synaptopodin in mouse renal cortex (scale bar = 40 μm). (B) Representative immunochemistry staining of PCSK9 in kidney tissue sections and statistical analyses (n = 6, scale bar = 20 μm). (C) Western blot and statistical analyses of PCSK9 protein expression in mouse renal cortex. β-Actin was used as loading control (n = 6). (D) Western blot and statistical analyses of KIM1 and NGAL protein expression in mouse renal cortex (n = 6). (E) Scr and (F) BUN levels in mouse plasma (n = 6). (G) Representative images of PAS staining in kidney tissue sections and tubular injury score (n = 6, ×200, scale bar = 50 μm; ×400, scale bar = 20 μm) Data are represented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The membranes were blocked with 5 % defatted milk for 1 h and then incubated with primary antibodies overnight at 4 ◦C: anti-PCSK9 (1:1000, 27882-1-AP, Proteintech, Wuhan, China), anti- kidney injury molecule 1 (KIM1) (1:1000, bs-2713R, Bioss, China), anti-neutrophil gelatinase-associated lipocalin (NGAL) (1:1000, 26991-1-AP, Proteintech, Wuhan, China), anti-megalin (1:200, sc-515772, Santa Cruz Biotechnology, USA), and anti-CAP1 (1:500, sc-376286, Santa Cruz Biotechnology, USA), anti-β-actin (1:8000, 20536-1-AP, Proteintech, Wuhan, China).

Techniques: Immunofluorescence, Staining, Western Blot, Expressing, Control, Clinical Proteomics

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Proprotein convertase subtilisin/kexin type 9 contributes to cisplatin-induced acute kidney injury by interacting with cyclase-associated protein 1 to promote megalin lysosomal degradation.

doi: 10.1016/j.bbamcr.2025.119984

Figure Lengend Snippet: Fig. 2. PCSK9 overexpression induces tubular injury and megalin downregulation. (A) 8-weeks-old C57BL/6J mice were injected with adeno-associated virus-human PCSK9 (AAV-PCSK9) via tail vein, two weeks after AAV injection, evolocumab (10 mg/kg) was injected subcutaneously, mice were killed at 12 weeks of age. (B) Western blot and statistical analyses of PCSK9 protein expression in mouse renal cortex (n = 3). (C) Representative immunochemistry staining of PCSK9 in kidney tissue sections and statistical analyses (n = 6, scale bar = 20 μm). (D) Representative immunochemistry staining of megalin in kidney tissue sections and statistical analyses (n = 6, scale bar = 20 μm). (E) Western blot and statistical analyses of megalin protein expression in mouse renal cortex (n = 6). (F) Western blot and statistical analyses of KIM1 and NGAL protein expression in mouse renal cortex (n = 6). (G) Western blot and statistical analyses of PCSK9 protein expression in HK2 cells (n = 4). (H) Western blot and statistical analyses of megalin protein expression in HK2 cells (n = 4). (I) Representative confocal images of BSA-FITC uptake in HK2 cells (scale bar = 25 μm). Data are represented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The membranes were blocked with 5 % defatted milk for 1 h and then incubated with primary antibodies overnight at 4 ◦C: anti-PCSK9 (1:1000, 27882-1-AP, Proteintech, Wuhan, China), anti- kidney injury molecule 1 (KIM1) (1:1000, bs-2713R, Bioss, China), anti-neutrophil gelatinase-associated lipocalin (NGAL) (1:1000, 26991-1-AP, Proteintech, Wuhan, China), anti-megalin (1:200, sc-515772, Santa Cruz Biotechnology, USA), and anti-CAP1 (1:500, sc-376286, Santa Cruz Biotechnology, USA), anti-β-actin (1:8000, 20536-1-AP, Proteintech, Wuhan, China).

Techniques: Over Expression, Injection, Virus, Western Blot, Expressing, Staining

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Proprotein convertase subtilisin/kexin type 9 contributes to cisplatin-induced acute kidney injury by interacting with cyclase-associated protein 1 to promote megalin lysosomal degradation.

doi: 10.1016/j.bbamcr.2025.119984

Figure Lengend Snippet: Fig. 3. PCSK9 inhibitor protects against cisplatin-induced kidney tubule injury. (A) Evolocumab (10 mg/kg) was injected into 8-weeks-old C57BL/6J mice sub cutaneously, 4 h later, cisplatin (25 mg/kg) was intraperitoneally injected, mice were killed 48 h after cisplatin injection. (B) Western blot and statistical analyses of PCSK9 protein expression in mouse renal cortex (n = 4). (C) Western blot and statistical analyses of KIM1 and NGAL protein expression in mouse renal cortex (n = 4). (D) Scr and (E) BUN levels in mouse plasma (n = 4). (F) Urinary albumin/creatinine in mouse urine (n = 4). (G) Representative images of PAS staining in kidney tissue sections and tubular injury score (n = 4, ×200, scale bar = 50 μm; ×400, scale bar = 20 μm). (H) Intrarenal lipid accumulation was evaluated by Oil red O (ORO) staining in kidney tissue sections (n = 4, scale bar = 20 μm). Data are represented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The membranes were blocked with 5 % defatted milk for 1 h and then incubated with primary antibodies overnight at 4 ◦C: anti-PCSK9 (1:1000, 27882-1-AP, Proteintech, Wuhan, China), anti- kidney injury molecule 1 (KIM1) (1:1000, bs-2713R, Bioss, China), anti-neutrophil gelatinase-associated lipocalin (NGAL) (1:1000, 26991-1-AP, Proteintech, Wuhan, China), anti-megalin (1:200, sc-515772, Santa Cruz Biotechnology, USA), and anti-CAP1 (1:500, sc-376286, Santa Cruz Biotechnology, USA), anti-β-actin (1:8000, 20536-1-AP, Proteintech, Wuhan, China).

Techniques: Injection, Western Blot, Expressing, Clinical Proteomics, Staining

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Proprotein convertase subtilisin/kexin type 9 contributes to cisplatin-induced acute kidney injury by interacting with cyclase-associated protein 1 to promote megalin lysosomal degradation.

doi: 10.1016/j.bbamcr.2025.119984

Figure Lengend Snippet: Fig. 4. PCSK9 inhibitor protects against megalin downregulation. (A) Western blot and statistical analyses of megalin protein expression in mouse renal cortex (n = 4). (B) Coomassie blue staining of polyacrylamide gels after urine protein electrophoresis. (C) Western blot and statistical analyses of KIM1 and NGAL protein expression in HK2 cells (n = 4). (D) Western blot and statistical analyses of megalin protein expression in HK2 cells (n = 4). (E) Representative confocal images of FITC-BSA uptake in HK2 cells (scale bar = 25 μm). Data are represented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The membranes were blocked with 5 % defatted milk for 1 h and then incubated with primary antibodies overnight at 4 ◦C: anti-PCSK9 (1:1000, 27882-1-AP, Proteintech, Wuhan, China), anti- kidney injury molecule 1 (KIM1) (1:1000, bs-2713R, Bioss, China), anti-neutrophil gelatinase-associated lipocalin (NGAL) (1:1000, 26991-1-AP, Proteintech, Wuhan, China), anti-megalin (1:200, sc-515772, Santa Cruz Biotechnology, USA), and anti-CAP1 (1:500, sc-376286, Santa Cruz Biotechnology, USA), anti-β-actin (1:8000, 20536-1-AP, Proteintech, Wuhan, China).

Techniques: Western Blot, Expressing, Staining, Protein Electrophoresis

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Proprotein convertase subtilisin/kexin type 9 contributes to cisplatin-induced acute kidney injury by interacting with cyclase-associated protein 1 to promote megalin lysosomal degradation.

doi: 10.1016/j.bbamcr.2025.119984

Figure Lengend Snippet: Fig. 6. Cisplatin enhances the binding of PCSK9 and CAP1, induce megalin downregulation and endocytosis dysfunction. (A) PCSK9 mRNA expression measured by qRT-PCR in HK2 cells (n = 6). (B) CAP1 mRNA expression measured by qRT-PCR in HK2 cells (n = 6). (C) Western blot and statistical analyses of PCSK9 and CAP1 protein expression in HK2 cells culture medium (n = 3). (D) Co-immunoprecipitation of PCSK9 from HK2 cells lysate. (E) Western blot and statistical analyses of KIM1 and NGAL protein expression in HK2 cells (n = 4). (F) Western blot and statistical analyses of megalin protein expression in HK2 cells (n = 4). (G) Repre sentative confocal images of FITC-BSA uptake in HK2 cells (scale bar = 25 μm). Data are represented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The membranes were blocked with 5 % defatted milk for 1 h and then incubated with primary antibodies overnight at 4 ◦C: anti-PCSK9 (1:1000, 27882-1-AP, Proteintech, Wuhan, China), anti- kidney injury molecule 1 (KIM1) (1:1000, bs-2713R, Bioss, China), anti-neutrophil gelatinase-associated lipocalin (NGAL) (1:1000, 26991-1-AP, Proteintech, Wuhan, China), anti-megalin (1:200, sc-515772, Santa Cruz Biotechnology, USA), and anti-CAP1 (1:500, sc-376286, Santa Cruz Biotechnology, USA), anti-β-actin (1:8000, 20536-1-AP, Proteintech, Wuhan, China).

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Western Blot, Immunoprecipitation

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Proprotein convertase subtilisin/kexin type 9 contributes to cisplatin-induced acute kidney injury by interacting with cyclase-associated protein 1 to promote megalin lysosomal degradation.

doi: 10.1016/j.bbamcr.2025.119984

Figure Lengend Snippet: Fig. 7. CAP1 knockout increases megalin levels in mouse overexpressing PCSK9 or in a mouse model of cisplatin-induced AKI. (A) CAP1 mRNA expression measured by qRT-PCR in mouse renal cortex (n = 6). (B) Western blot and statistical analyses of CAP1 protein expression in mouse renal cortex (n = 6). (C, D) Western blot and statistical analyses of megalin protein expression in mouse renal cortex (n = 4). (E) Coomassie blue staining of polyacrylamide gels after urine protein electro phoresis. (F) Western blot and statistical analyses of KIM1 and NGAL protein expression in mouse renal cortex (n = 4). (G) Scr and (H) BUN levels in mouse plasma (n = 4). (I) Representative images of PAS staining in kidney tissue sections and tubular injury score (n = 4, ×200, scale bar = 50 μm; ×400, scale bar = 20 μm). Data are represented as the mean ± SD, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The membranes were blocked with 5 % defatted milk for 1 h and then incubated with primary antibodies overnight at 4 ◦C: anti-PCSK9 (1:1000, 27882-1-AP, Proteintech, Wuhan, China), anti- kidney injury molecule 1 (KIM1) (1:1000, bs-2713R, Bioss, China), anti-neutrophil gelatinase-associated lipocalin (NGAL) (1:1000, 26991-1-AP, Proteintech, Wuhan, China), anti-megalin (1:200, sc-515772, Santa Cruz Biotechnology, USA), and anti-CAP1 (1:500, sc-376286, Santa Cruz Biotechnology, USA), anti-β-actin (1:8000, 20536-1-AP, Proteintech, Wuhan, China).

Techniques: Knock-Out, Expressing, Quantitative RT-PCR, Western Blot, Staining, Clinical Proteomics

Journal: The FASEB Journal

Article Title: Novel inflammatory markers in intracerebral hemorrhage: Results from Olink proteomics analysis

doi: 10.1096/fj.202402183rr

Figure Lengend Snippet: FIGURE 6 The renal function impairment in the two groups of patients and its correlation with KIM1. (A) Serum biochemical parameters of creatinine (Cr), blood Urea Nitrogen (BUN), uric acid (UA), and electrolytes in the two groups of patients. (B) Heatmap of the correlation between KIM1 and Cr, BUN, UA, and electrolytes. *p < .05, ***p < .001.

Article Snippet: The ELISA kits for the first four inflammatory factors are: Human Kidney Injury Molecule 1 (KIM1) (E- EL- H6029, Elabscience, Wuhan, China), Human Interleukin 6 (IL6) (E- EL- H6156, Elabscience, Wuhan, China), Human pentraxin3 (PTX3) (E- EL- H6081, Elabscience, Wuhan, China), and Human Galection- 9 (Gal- 9) (KE00175, Proteintech, Rosemont, USA).

Techniques: