Journal: Experimental and Therapeutic Medicine

Article Title: Reduced expression of microRNA-199a-3p is associated with vascular endothelial cell injury induced by type 2 diabetes mellitus

doi: 10.3892/etm.2018.6655

Figure Lengend Snippet: Effect of miR-199a-3p overexpression on the autophagy of HUVECs. (A) The number of autophagosomes in HUVECs was determined using laser-scanning confocal microscopy (magnification, ×200). (B) Western blotting was performed to assess the relative expression of autophagy-associated LC3BII protein in HUVECs *P<0.05 vs. NC. NC, negative control; miR, microRNA; HUVEC, human umbilical vein endothelial cell; LC3BII.

Article Snippet: Protein samples (10 µg) were mixed with 5X SDS loading buffer and the mixture was denatured by boiling in a water bath for 10 min. Proteins were separated by 10% SDS-PAGE and transferred to polyvinylidene difluoride membranes, which were subsequently blocked with 5% skimmed milk at room temperature for 1 h. Membranes were incubated with phosphatidylinositol 3-kinase (PI3K) catalytic subunit p110α (1:1,000; cat. no. 4249; Cell Signaling Technology, Inc., Danvers, MA, USA), PI3K regulatory subunit p85 (1:1,000; cat. no. AF1729; Beyotime Institute of Biotechnology) LC3BII (1:1,000; cat. no. AL221; Beyotime Institute of Biotechnology) and mouse anti-human GAPDH (1:5,000; cat. no. AF0006; Beyotime Institute of Biotechnology) primary antibodies at 4°C overnight.

Techniques: Over Expression, Confocal Microscopy, Western Blot, Expressing, Negative Control

Journal: PLoS ONE

Article Title: Prolyl Oligopeptidase Inhibition Attenuates Steatosis in the L02 Human Liver Cell Line

doi: 10.1371/journal.pone.0165224

Figure Lengend Snippet: (A-F) Oil red O staining of control cells (A), FFA-treated cells (B), cells treated with FFA+ S17902 (0.026 μM), (C) FFA+ S17902 (0.26 μM), (D) FFA+ S17902 (13 μM), (E) or FFA+ S17902 (26 μM) (F) for 24 h; (G) TG concentration in the L02 cells treated with S17092 for 24 h.(H) FASN/PPAR-γ/SREBP-1c mRNA expression levels in the controls and FFA-treated L02 cells with or without S17092 incubation for 24 h. (I-J) LC3B II protein expression levels in the controls and FFA-treated L02 cells with or without S17092 incubation for 24 h. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Indeed, our results demonstrated that LC3B II, a bioprotein marker of autophagy, was significantly increased in the L02 cells that were treated with FFA for 24 h, which implies enhanced autophagy and is in agreement with a previous study [ ].

Techniques: Staining, Control, Concentration Assay, Expressing, Incubation