Journal: Acta Cirúrgica Brasileira

Article Title: MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model ¹

doi: 10.1590/s0102-865020190060000002

Figure Lengend Snippet: MiR-106a was up-regulated in serum of sepsis patients and CLP-induced mice models. The relative miR-106a level was tested by qRT-PCR in the serum from sepsis and healthy patients, CLP-induced sepsis mouse models and TCMK-1 cells treated with LPS. *** p<0.001.

Article Snippet: TCMK-1 mouse kidney epithelial cell line was purchased from BioVector NTCC Inc. (Beijing, China) and cultured in 90% high-glucose Dulbecco’s modified Eagle medium (DMEM, Solarbio, Beijing, China) and 10% fetal bovine serum (FBS, Solarbio, Beijing, China) at 37°C.

Techniques: Quantitative RT-PCR

Journal: Acta Cirúrgica Brasileira

Article Title: MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model ¹

doi: 10.1590/s0102-865020190060000002

Figure Lengend Snippet: Knockdown of miR-106a reduced LPS-induced apoptosis in TCMK-1 cells. miR-106a inhibitor, NC inhibitor and LPS were transfected into TCMK-1 cells. (A) Relative miR-106a level, (B) cell viability, (C) relative caspase-3 activity and (D) the protein level of Bax, cleaved caspase-3 and Bcl-2 were detected by qRT-PCR, CCK-8 and western blotting. ** p<0.01; *** p<0.001.

Article Snippet: TCMK-1 mouse kidney epithelial cell line was purchased from BioVector NTCC Inc. (Beijing, China) and cultured in 90% high-glucose Dulbecco’s modified Eagle medium (DMEM, Solarbio, Beijing, China) and 10% fetal bovine serum (FBS, Solarbio, Beijing, China) at 37°C.

Techniques: Transfection, Activity Assay, Quantitative RT-PCR, CCK-8 Assay, Western Blot

Journal: Acta Cirúrgica Brasileira

Article Title: MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model ¹

doi: 10.1590/s0102-865020190060000002

Figure Lengend Snippet: Knockdown of miR-106a reduced LPS-induced inflammatory factor levels in TCMK-1 cells. miR-106a inhibitor, NC inhibitor and LPS were transfected into TCMK-1 cells. (A) The relative mRNA level and (B) content of TNF-α, IL-1β, IL-6 were detected by qRT-PCR and ELISA. * p<0.05; ** p<0.01; *** p<0.001.

Article Snippet: TCMK-1 mouse kidney epithelial cell line was purchased from BioVector NTCC Inc. (Beijing, China) and cultured in 90% high-glucose Dulbecco’s modified Eagle medium (DMEM, Solarbio, Beijing, China) and 10% fetal bovine serum (FBS, Solarbio, Beijing, China) at 37°C.

Techniques: Transfection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Acta Cirúrgica Brasileira

Article Title: MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model ¹

doi: 10.1590/s0102-865020190060000002

Figure Lengend Snippet: THBS2 was a target of miR-106a. MiR-106a inhibitor, NC inhibitor, and miR-106a mimics, NC mimics, THBS2-WT and THBS2-MUT were transfected into HEK293 and TCMK-1 cells. (A) The binding site was predicted by Targetscan website. (B) Relative miR-106a level was tested by qRT-PCR. (C) The target relationship between miR-106a and THBS2 was confirmed by dual luciferase reporter assay. (D) THBS2 expression was detected by western blotting. ** p<0.01; *** p<0.001.

Article Snippet: TCMK-1 mouse kidney epithelial cell line was purchased from BioVector NTCC Inc. (Beijing, China) and cultured in 90% high-glucose Dulbecco’s modified Eagle medium (DMEM, Solarbio, Beijing, China) and 10% fetal bovine serum (FBS, Solarbio, Beijing, China) at 37°C.

Techniques: Transfection, Binding Assay, Quantitative RT-PCR, Luciferase, Reporter Assay, Expressing, Western Blot

Journal: Acta Cirúrgica Brasileira

Article Title: MiR-106a aggravates sepsis-induced acute kidney injury by targeting THBS2 in mice model ¹

doi: 10.1590/s0102-865020190060000002

Figure Lengend Snippet: MiR-106a increased LPS-induced inflammation and apoptosis of TCMK-1 cells via regulating THBS2 expression. MiR-106a inhibitor, NC inhibitor, and miR-106a mimics, NC mimics, siTHBS2 and siNC were transfected into LPS-treated TCMK-1 cells. (A) Relative THBS2 mRNA level, (B) cell viability, (C) relative caspase-3 activity, (D-E) the relative mRNA level and content of TNF-α, IL-1β, IL-6, as well as (F) the protein level of Bax, cleaved caspase-3 and Bcl-2 were detected by qRT-PCR, CCK-8, ELISA and western blotting. * p<0.05; ** p<0.01; *** p<0.001.

Article Snippet: TCMK-1 mouse kidney epithelial cell line was purchased from BioVector NTCC Inc. (Beijing, China) and cultured in 90% high-glucose Dulbecco’s modified Eagle medium (DMEM, Solarbio, Beijing, China) and 10% fetal bovine serum (FBS, Solarbio, Beijing, China) at 37°C.

Techniques: Expressing, Transfection, Activity Assay, Quantitative RT-PCR, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Western Blot