Journal: Antioxidants

Article Title: YAK577 Attenuates Vascular Calcification by Targeting an MMP14–NOX2/ROS Axis in VSMCs and a Vitamin D 3 -Induced Mouse Model

doi: 10.3390/antiox15050605

Figure Lengend Snippet: The HDAC inhibitor YAK577 reduces CM-induced vascular calcification in VSMCs. ( A ) Chemical structure of YAK577. ( B ) Effect of YAK577 on VSMC viability determined by MTT assay ( n = 4, 5, 5, 5, 4). ( C ) Representative images of Alizarin Red S-stained VSMCs observed under a light microscope. Scale bar = 200 μm. Bottom: Macroscopic photograph of Alizarin Red S-stained VSMCs captured using a smartphone camera, showing red coloration corresponding to calcium deposition. ( D ) Quantitative analysis of calcium content in CM-treated VSMCs with or without YAK577 treatment ( n = 3, 3, 3). Data are presented as mean ± SEM. *** p < 0.001; NS, not significant. GM, growth medium; CM, calcification medium. Each dot represents one independent biological experiment (not a technical replicate).

Article Snippet: The rat A10 vascular smooth muscle cell line (ATCC CRL-1476) was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: MTT Assay, Staining, Light Microscopy

Journal: Antioxidants

Article Title: YAK577 Attenuates Vascular Calcification by Targeting an MMP14–NOX2/ROS Axis in VSMCs and a Vitamin D 3 -Induced Mouse Model

doi: 10.3390/antiox15050605

Figure Lengend Snippet: MMP14 overexpression promotes NOX2 upregulation and enhances osteogenic marker expression in A10 cells. ( A – D ) qRT-PCR analysis of Mmp14 ( n = 8, 8), Bmp2 ( n = 10, 8), Runx2 ( n = 10, 8), and Msx2 ( n = 12, 12) mRNA levels in A10 cells following MMP14 plasmid transfection. ( E , F ) Representative Western blots and corresponding densitometric quantification showing increased expression of MMP14 ( n = 7, 7), RUNX2 ( n = 9, 9), BMP2 ( n = 8, 8) and NOX2 ( n = 7, 7) proteins after MMP14 overexpression. β-actin was used as a loading control. ( G ) Representative immunofluorescence images of A10 cells transfected with vector or GFP-MMP14. GFP-MMP14 was detected with anti-GFP antibody/Alexa Fluor 488 (green), F-actin with Texas Red-X Phalloidin (red), and nuclei with DAPI (blue). Scale bar = 100 μm. Data are presented as mean ± SEM. *** p < 0.001; ** p < 0.01; NS, not significant. Each dot represents one independent biological experiment (not a technical replicate).

Article Snippet: The rat A10 vascular smooth muscle cell line (ATCC CRL-1476) was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Over Expression, Marker, Expressing, Quantitative RT-PCR, Plasmid Preparation, Transfection, Western Blot, Control, Immunofluorescence