Journal: Cell Death & Disease

Article Title: Proximal tubular RAGE mediated the renal fibrosis in UUO model mice via upregulation of autophagy

doi: 10.1038/s41419-022-04856-z

Figure Lengend Snippet: A – C Immunoblot analysis of RAGE and β-Tubulin in BUMPT cells treated with or without TGF-β1 and the kidneys of UUO mice and Ob patients. D – F Analysis of the grayscale image between them. G Immunohistochemical staining of RAGE in the kidney of the Sham or UUO mice. Original magnification×400. Scale bar: 100 µM. Data are expressed as means ± s.d. ( n = 6). # P < 0.05 versus 0 d & 0 h and Con group. Each experiment A – C was repeated six times independently with similar results. B – F indicate the statistical Student’s t test used (means ± s.d., n = 6, P < 0.05).

Article Snippet: BUMPT cells were grown in DMEM medium (Thermo-Fisher-Scientific) containing 10% FBS and antibiotics in a 37 °C incubator with 5% CO 2 .

Techniques: Western Blot, Immunohistochemical staining, Staining

Journal: Cell Death & Disease

Article Title: Proximal tubular RAGE mediated the renal fibrosis in UUO model mice via upregulation of autophagy

doi: 10.1038/s41419-022-04856-z

Figure Lengend Snippet: The plasmid and siRNA of RAGE were transfected into BUMPT cells and treated with or without 5 ng/ml TGF-β1 for 24 h. A , G Immunoblot analysis of fibronectin & Col I&III, vimentin, and RAGE. B – F , H – L Analysis of the gray scale image between them. Data are expressed as means ± s.d. ( n = 6). # P < 0.05 versus NC or Vector with the control group. * P < 0.05 versus NC or Vector with the TGF-β1 group. Each experiment ( A , G ) was repeated six times independently with similar results. B – F , H – L indicate the statistical Student’s t test used (means ± s.d., n = 6, P < 0.05).

Article Snippet: BUMPT cells were grown in DMEM medium (Thermo-Fisher-Scientific) containing 10% FBS and antibiotics in a 37 °C incubator with 5% CO 2 .

Techniques: Plasmid Preparation, Transfection, Western Blot, Control

Journal: Cell Death & Disease

Article Title: Proximal tubular RAGE mediated the renal fibrosis in UUO model mice via upregulation of autophagy

doi: 10.1038/s41419-022-04856-z

Figure Lengend Snippet: The plasmid and siRNA of RAGE were transfected into BUMPT cells or HK-2 cells and treated with or without 5 ng/ml TGF-β1 for 24 h. A , B Immunoblot analysis of LC3 & p62 & RAGE in BUMPT cells. C , D Analysis of the grayscale image between them. # P < 0.05 versus NC or Vector with the control group. * P < 0.05 versus NC or Vector with the TGF-β1 group. E After GFP-LC3 transfection, HK-2 cells were transfected with or without the plasmid or siRNA of RAGE, followed by the treatment with TGF-β1 for 24 h. The punctate accumulation of GFP-LC3 was visualized under fluorescence microscopy. F And the average number of GFP-LC3 dots per cell was then calculated. # P < 0.05 versus NC with the control group. ^ P < 0.05 versus NC with the TGF-β1 group. * P < 0.05 versus NC with the TGF-β1 group. Each experiment A , B , E was repeated six times independently with similar results. C , D , F indicate the statistical Student’s t test used (means ± s.d., n = 6, P < 0.05).

Article Snippet: BUMPT cells were grown in DMEM medium (Thermo-Fisher-Scientific) containing 10% FBS and antibiotics in a 37 °C incubator with 5% CO 2 .

Techniques: Plasmid Preparation, Transfection, Western Blot, Control, Fluorescence, Microscopy

Journal: Cell Death & Disease

Article Title: Proximal tubular RAGE mediated the renal fibrosis in UUO model mice via upregulation of autophagy

doi: 10.1038/s41419-022-04856-z

Figure Lengend Snippet: Immunoblotting results demonstrated that TGF-β1-induced the increasing of p-Stat3, Atg7, and LC3 II, and the reduction of p62 was notably attenuated by STAT3-IN-1, an STAT3 inhibitor. A Immunoblot analysis of LC3 & p62 & Atg7 & p-Stat3 & Stat3 in BUMPT cells. B Analysis of the grayscale image between them. # P < 0.05 versus Saline with the control group. * P < 0.05 versus Saline with the TGF-β1 group. C After GFP-LC3 transfection, HK-2 cells were transfected with or without STAT3-IN-1, followed by the treatment with TGF-β1 for 24 h. The punctate accumulation of GFP-LC3 was visualized under fluorescence microscopy. D The number of LC3 puncta per cell was calculated. # P < 0.05 versus Saline with the control group. * P < 0.05 versus Saline with the TGF-β1 group. E ChIP assays verified that a binding sites (a 227 bp fragment) of STAT3 existed in the promoter region of Atg7. The plasmid and siRNA of RAGE were transfected into BUMPT cells and treated with or without 5 ng/ml TGF-β1 for 24 h. F , G Immunoblot analysis of Atg7 & p-Stat3 & Stat3 in BUMPT cells. H , I Analysis of the grayscale image between them. # P < 0.05 versus NC or Vector with the control group. * P < 0.05 versus Saline with the TGF-β1 group. Each experiment A , C , F , G was repeated six times independently with similar results. B , H , I indicate the statistical Student’s t test used (means ± s.d., n = 6, P < 0.05).

Article Snippet: BUMPT cells were grown in DMEM medium (Thermo-Fisher-Scientific) containing 10% FBS and antibiotics in a 37 °C incubator with 5% CO 2 .

Techniques: Western Blot, Saline, Control, Transfection, Fluorescence, Microscopy, Binding Assay, Plasmid Preparation

Journal: Cell Death & Disease

Article Title: Proximal tubular RAGE mediated the renal fibrosis in UUO model mice via upregulation of autophagy

doi: 10.1038/s41419-022-04856-z

Figure Lengend Snippet: The plasmid and siRNA of Atg7 were transfected into BUMPT cells and treated with or without 5 ng/ml TGF-β1 for 24 h. A , C Immunoblot analysis of Atg7 & LC3 & p62 & FN & Collagen I & Collagen III and Vimentin in BUMPT cells. B , D Analysis of the grayscale image between them. # P < 0.05 versus NC or Vector with the control group. * P < 0.05 versus NC or Vector with the TGF-β group. E After GFP-LC3 transfection, HK-2 cells were transfected with or without the plasmid or siRNA of ATG7, followed by the treatment with TGF-β1 for 24 h. The punctate accumulation of GFP-LC3 was visualized under fluorescence microscopy. F The number of LC3 puncta per cell was calculated. # P < 0.05 versus NC or Vector with the control group. ^ P < 0.05 versus NC or Vector with the TGF-β group. * P < 0.05 versus NC or Vector with the TGF-β group. Each experiment A , C , E was repeated six times independently with similar results. B , D , F indicate the statistical Student’s t test used (means ± s.d., n = 6, P < 0.05).

Article Snippet: BUMPT cells were grown in DMEM medium (Thermo-Fisher-Scientific) containing 10% FBS and antibiotics in a 37 °C incubator with 5% CO 2 .

Techniques: Plasmid Preparation, Transfection, Western Blot, Control, Fluorescence, Microscopy