Journal: Acta Biochimica et Biophysica Sinica

Article Title: Circular RNA-VPS13A attenuates diabetes-induced enteric glia damage by targeting miR-182/GDNF axis

doi: 10.3724/abbs.2022073

Figure Lengend Snippet: The role of GDNF in the regulatory effect on circVPS13A-miR-182 on EGC damage in DM (A) The predicted binding site for miR-182 in the 3’-UTR of GDNF. (B) The luciferase reporter vector containing WT GDNF 3’-UTR or MUT GDNF 3’-UTR and miR-NC or miR-182 mimic were co-transfected into HK293T cells, and luciferase activity was determined and normalized to Renilla luciferase activity. (C) The effect of transfection with miR-182 mimic on GDNF expression in CRL-2690 cells was assessed by qPCR, western blot analysis, and ELISA. (D) The effect of miR-182 inhibitor or circVPS13A overexpression on GDNF expression in HG-treated CRL-2690 cells by qPCR, western blot analysis, and ELISA. (E) The correlation analysis of GDNF with miR-182 or circVPS13A in CRL-2690 cells with or without HG treatment. ** P<0.01; ## P<0.01.

Article Snippet: To assess the effect of miR-182 on GDNF, HEK293 cells were co-transfected with 100 pM of either miR-182 mimic or control, 40 ng of either pGL3-GDNF-3′UTR or pGL3-GDNF-3′UTR MUT (Genepharma), and 4 ng of pRL-TK (Promega) using Lipofectamine 2000 (Life Technologies).

Techniques: Binding Assay, Luciferase, Plasmid Preparation, Transfection, Activity Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Over Expression

Journal: Oncotarget

Article Title: A novel pan-Nox inhibitor, APX-115, protects kidney injury in streptozotocin-induced diabetic mice: possible role of peroxisomal and mitochondrial biogenesis

doi: 10.18632/oncotarget.18540

Figure Lengend Snippet: Diabetes was induced in mice by intraperitoneal injection of STZ (50 mg/kg). Then APX-115 (60 mg/kg/day) or losartan (1.5 mg/kg/day) was administered orally for 12 weeks to diabetic mice. After 12 weeks, urine and blood samples were collected for analysis of ( A ) urinary albumin excretion, ( B ) albumin/creatinine ratio, ( C ) creatinine clearance rate, and ( D ) plasma cystatin C. ( E ) Kidneys were fixed in paraffin and cut into 3 μm sections that were subsequently stained with PAS reagent. Scale bar: 10 μm; original magnification: 630×. After PAS staining, ( F ) glomerular volume, ( G ) mesangial area, and ( H ) tuft area were analyzed using Image-Pro Plus 4.5.1. DM, STZ-induced diabetic mice. Data are presented as means ± SE of 10–12 mice/group; * p < 0.05 vs. control, † p < 0.05 vs. DM.

Article Snippet: A murine mesangial cell line (MES-13, cloned from mice transgenic for the early region of SV-40 virus, passage 25) was obtained from American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Injection, Clinical Proteomics, Staining, Control

Journal: Oncotarget

Article Title: A novel pan-Nox inhibitor, APX-115, protects kidney injury in streptozotocin-induced diabetic mice: possible role of peroxisomal and mitochondrial biogenesis

doi: 10.18632/oncotarget.18540

Figure Lengend Snippet: ( A ) Plasma LPO, ( B ) urinary LPO, ( C ) kidney tissue LPO, ( D ) Nox1, ( E ) Nox2, and ( F ) Nox4 mRNA expression levels in kidneys were measured using real-time PCR. ( G and H ) Frozen kidney sections were stained with DHE at 5 µM (original magnification: 400×; scale bar: 20 μm). (A–H) Data are presented as means ± SE of 10–12 mice/group; * p < 0.05 vs. control, † p < 0.05 vs. DM. ( I ) Mesangial cells were incubated with or without APX-115 (1 µM) for 30 min and stimulated with or without 30 mM high glucose (HG) for 24 h followed by angII for 30 min. After that cells were incubated with 10 µM DCF-DA for 10 min and the fluorescence intensity was measured with a Zeiss vision system. Data are presented as means ± SE of at least 2 independent experiments; * p < 0.05 vs. control, † p < 0.05 vs. angII or angII+HG in DMSO.

Article Snippet: A murine mesangial cell line (MES-13, cloned from mice transgenic for the early region of SV-40 virus, passage 25) was obtained from American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Clinical Proteomics, Expressing, Real-time Polymerase Chain Reaction, Staining, Control, Incubation, Fluorescence