Journal: International Journal of Molecular Medicine

Article Title: Long non-coding RNA AL355711 promotes smooth muscle cell migration through the ABCG1/MMP3 pathway

doi: 10.3892/ijmm.2021.5040

Figure Lengend Snippet: ABCG1 and MMP3 are highly expressed in an animal model of atherosclerosis. An animal model of atherosclerosis was successfully constructed by feeding 6-week-old apolipoprotein E knockout mice with a Western HFD for 12 weeks. (A) Representative en face images of the ascending aorta stained with Oil Red O (×2.5 magnification). A blue mouse pad was used as the supporting platform for the ascending aortas (hence the green-like background). Data shown in the graphs are mean ± SD. * P<0.05. In total, 4 mice were used as the controls and 5 mice were used in the HFD group. The experiments were performed once. (B) Aortic root cross-sections stained with H&E. Scale bar left panel, 250 µ m (×100 magnification); scale bar right panel, 100 µ m (×200 magnification). The images on the right panel are an enlarge magnification of the red boxed areas of the images in the left panels. (C) Aortic root cross-sections stained with Masson's stain indicated that the HFD diet significantly reduced the collagen fiber content. Data shown in the graphs are the mean ± SD. * P<0.05. Scale bar left panel, 250 µ m (×100 magnification); scale bar right panel, 100 µ m (×200 magnification). The images on the right panel are an enlarge magnification of the red boxed areas of the images in the left panels. (D) ABCG1 immunohistochemical staining revealed that the HFD significantly increased the ABCG1 levels in plaques. Data shown in the graphs are the mean ± SD. * P<0.05. Scale bar left panel, 250 µ m (×100 magnification); scale bar right panel, 100 µ m (×200 magnification). The images on the right panel are an enlarge magnification of the red boxed areas of the images in the left panels. (E) MMP3 immunohistochemical staining revealed that the HFD significantly increased the level of ABCG1 in plaques. Data shown in the graphs are the mean ± SD. * P<0.05. Scale bar left panel, 250 µ m (×100 magnification); scale bar right panel, 100 µ m (×200 magnification). The images on the right panel are an enlarge magnification of the red boxed areas of the images in the left panels. (F) MMP3 immunohistochemical staining revealed that the HFD significantly increased the level of ABCG1 in plaques. Data shown in the graphs are the mean ± SD. * P<0.05. The same paraffin-embedded tissue sections were stained for MMP3 and ABCG1, both of which were highly expressed in the same section of atherosclerotic plaques. Scale bar left panel, 100 µ m (×200 magnification); arrows indicate the red boxed areas, which are presented as an enlarged magnification in the images on the right (×20,000 magnification). lncRNA, long non-coding RNA; ABCG1, ATP-binding cassette sub-family G member 1; MMP3, matrix metalloproteinase 3; HFD, high-fat diet; H&E, hematoxylin and eosin.

Article Snippet: The mouse aortic roots were fixed in 4% paraformaldehyde (Wuhan Servicebio Technology Co., Ltd.) 20 min at room temperature, and stained for 2 h using Oil Red O (O0625; Sigma-Aldrich; Merck KGaA), and Masson's stain or hematoxylin and eosin (H&E) staining according to the manufacturer's instructions (G1346 and G1120; Beijing Solarbio Science & Technology Co., Ltd.).

Techniques: Animal Model, Construct, Knock-Out, Western Blot, Staining, Immunohistochemical staining, Binding Assay