Journal: Cardiovascular Diabetology
Article Title: Lysolecithin reprogramming via LPCAT1 modulation restores endothelial function and prevents diabetes-associated dysmetabolism
doi: 10.1186/s12933-025-03053-4
Figure Lengend Snippet: LPCAT1 is involved in hyperglycemia-induced vascular damage and lysolecithin homeostasis. A Relative mRNA expression is given as relative quantification (RQ = 2 −ΔΔCT ) values of LPCAT1, LPCAT2, LPCAT3, and LPCAT4. 2 −ΔΔCT values of the first controls were normalized to 1. B Representative immunoblots and densitometric analyses of 3 independent experiments evaluating protein levels of LPCAT1 and LPCAT2. GAPDH has been used as internal loading control for normalization expression in HUVECs. C Immunofluorescence analysis of VE-cadherin and LPCAT1 signaling end relative co-localization plots in HUVECs. Scale bar: 25 μm. D Vascular reactivity assay, dose-response curves to acetylcholine (ACh) in ex vivo C57BL/6 mouse mesenteric arteries under basal condition (CTRL) after three hours of high glucose exposure (30 mM) and pretreated with SP6 for two hours before HG exposure. E Graphical representation of endothelial cells (ECs) and smooth muscle cells (SMCs) isolation from pooled mice mesenteric arteries exposed to different experimental conditions; F : Relative mRNA expression is given as relative quantification (RQ = 2 −ΔΔCT ) values of LPCAT1, LPCAT2, GLUT1, GLUT2, and GLUT4 into isolated ECs ( upper ) and in SMCs ( bottom ). 2 −ΔΔCT values of the first controls were normalized to 1
Article Snippet: The primary antibody used were anti-API5 (Abcam, UK; Cat# ab65836) diluted at 1:1000; anti-VEGF (Santa Cruz Biotechnology, USA; Cat# sc-7269) diluted at 1:500; anti- LPCAT2 (Proteintech Group, USA; Cat# 15082-1-AP) diluted at 1:5000; anti- LPCAT1 (Proteintech Group, USA; Cat# 16112-1-AP) diluted at 1:1000; anti-GLUT1 (Cohesion Biosciences, UK; Cat# CPA2071) diluted at 1:1000; anti-GLUT4 (ABclonal, DE; Cat# A7637) diluted at 1:1000; Anti-VE Cadherin (Abcam, UK; Cat# ab33168) diluted at 1:500.
Techniques: Expressing, Quantitative Proteomics, Western Blot, Control, Immunofluorescence, Ex Vivo, Isolation
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