Journal: International journal of molecular sciences
Article Title: Sestrin2 Suppression Promotes Endothelial-Mesenchymal Transition and Exacerbates Methylglyoxal-Induced Endothelial Dysfunction.
doi: 10.3390/ijms252413463
Figure Lengend Snippet: Figure 3. Effect of SESN2 silencing on the protein expression of endothelial and mesenchymal markers in endothelial cells subjected to MGO. (A) Western bot analysis of endothelial markers: VE-Cadherin, PECAM, and Tie2. (B–D) Densitometry data of protein expression of VE-Cadherin (B), PECAM (C), and Tie2 (D) normalized against loading control GAPDH and expressed as a percentage (%) of the untreated group (CTRL). (E) Western bot analysis of mesenchymal markers: α-SMA, Vimentin, and TGF-β. (F–H) Densitometry data of protein expression of α-SMA (F), Vimentin (G), and TGF-β (H) normalized against loading control GAPDH and expressed as a percentage (%) of the untreated group (CTRL). Cells were left untreated or incubated with SESN2 siRNA duplexes for 48 h, and then exposed to MGO (600 µM for 18 h). Data are presented as mean ± S.E.M (n = 4 in each group). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, vs. CTRL or indicated groups.
Article Snippet: Following blocking, membranes were washed with TBS-T and incubated overnight at 4 ◦C on a shaker with the following primary antibodies: SESN2 (#8487S), vascular endothelial (VE)-cadherin (#2500S), platelet endothelial cell adhesion molecule (PECAM-1) (#77699S), tyrosine kinase with immunoglobin and EGF homology domains 2 (Tie2) (#7403S), vimentin (#5741S), transforming growth factor (TGF)-β (#3711S), GAPDH (#2118S) (Cell Signalling Technology, Danvers, MA, USA), and alpha-smooth muscle actin (α-SMA) (#ab7817) (Abcam, Cambridge, UK).
Techniques: Expressing, Western Blot, Control, Incubation
