Journal: American Journal of Translational Research

Article Title: microRNA-1246 mediates lipopolysaccharide-induced pulmonary endothelial cell apoptosis and acute lung injury by targeting angiotensin-converting enzyme 2

doi:

Figure Lengend Snippet: LPS induces miR-1246 to target ACE2 expression in PMVECs. A: qRT-PCR analysis of miR-1246 expression in PMVECs treated with LPS or vehicle control. *P < 0.05 vs. vehicle. B: Western blot analysis of ACE2 protein in PMVECs transfected with miR-1246 mimic or negative control miRNA. Bar graphs (bottom) show densitometric analysis of ACE2 protein. *P < 0.05 vs. vehicle. C: Luciferase reporter assay showed that miR-1246 repressed the expression of the luciferase reporter containing the wild-type (wt) but not mutated (mut) ACE2 3’-UTR. The predicted target site for miR-1246 in the ACE2 3’-UTR is shown in top panels. *P < 0.05 vs. vehicle. D: Western blot analysis of ACE2 protein in PMVECs pretransfected with anti-miR-1246 or control inhibitors before exposure to LPS. *P < 0.05.

Article Snippet: Cell culture and LPS treatment Human PMVECs were purchased from ScienCell Research Laboratories, Inc. (Carlsbad, CA, USA) and cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS; GE Healthcare HyClone, Logan, UT, USA).

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

Journal: American Journal of Translational Research

Article Title: microRNA-1246 mediates lipopolysaccharide-induced pulmonary endothelial cell apoptosis and acute lung injury by targeting angiotensin-converting enzyme 2

doi:

Figure Lengend Snippet: miR-1246 is involved in LPS-triggered apoptosis and inflammatory response in PMVECs. A: TUNEL staining of PMVECs pretransfected with anti-miR-1246 or control inhibitors (C-inhibitor) before exposure to LPS. Scale bar, 50 μm. *P < 0.05. B: ELISA analysis of IL-1β and TNF-α levels in conditioned medium from PMVECs with indicated treatments. *P < 0.05. C: TUNEL staining of PMVECs transfected with miR-1246 mimic or negative control miRNA. Scale bar, 50 μm. D: Measurement of IL-1β and TNF-α levels in conditioned medium from PMVECs transfected with miR-1246 mimic or negative control miRNA. *P < 0.05 vs. control.

Article Snippet: Cell culture and LPS treatment Human PMVECs were purchased from ScienCell Research Laboratories, Inc. (Carlsbad, CA, USA) and cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS; GE Healthcare HyClone, Logan, UT, USA).

Techniques: TUNEL Assay, Staining, Control, Enzyme-linked Immunosorbent Assay, Transfection, Negative Control

Journal: American Journal of Translational Research

Article Title: microRNA-1246 mediates lipopolysaccharide-induced pulmonary endothelial cell apoptosis and acute lung injury by targeting angiotensin-converting enzyme 2

doi:

Figure Lengend Snippet: miR-1246 promotes PMVEC apoptosis by repressing ACE2 expression. A: Western blot analysis of ACE2 protein in PMVECs transfected with indicated constructs. B: Detection of apoptosis by TUNEL staining in PMVECs transfected with indicated constructs. *P < 0.05. C: Western blot analysis of Bcl-2, Bcl-xL, and Bax proteins in PMVECs transfected with indicated constructs.

Article Snippet: Cell culture and LPS treatment Human PMVECs were purchased from ScienCell Research Laboratories, Inc. (Carlsbad, CA, USA) and cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS; GE Healthcare HyClone, Logan, UT, USA).

Techniques: Expressing, Western Blot, Transfection, Construct, TUNEL Assay, Staining