Morphological changes in organelles under atherosclerotic stimulations with high glucose (HG) and oxidized low-density lipoprotein (ox-LDL) are associated with cellular stress responses. (A) Representative image of ER labeled with GFP-Sec61β in human aortic VSMCs (HA-VSMCs) and human carotid VSMCs (HC-VSMCs) treated with dimethyl sulfoxide (DMSO; control group, Ctrl.), dithiothreitol (DTT; 3 mM), HG (20 mM), and ox-LDL (50 μg/mL) for 24 h. (B) Quantitative analysis of the percentage of cells with ER whorls in HA-VSMCs and HC-VSMCs treated with DMSO, DTT, HG, and ox-LDL (n = 30 cells in each group). (C) Western blotting analysis of PERK, spliced X-box-binding protein 1 (XBP1-s) and unspliced XBP1 (XBP1-u) in HA-VSMCs and HC-VSMCs treated with DMSO, DTT, HG, and ox-LDL (n = 3 times each group). (D) Representative image of mitochondria stained with deep red Mitotracker in HA-VSMCs and HC-VSMCs treated with DMSO (Ctrl.), oligomycin A1 (Oligo, 1 μM), HG (20 mM), and ox-LDL (50 μg/mL) for 24 h. (E) Quantitative analysis of the aspect ratio in HA-VSMCs and HC-VSMCs treated with DMSO, Oligo, HG, and ox-LDL (n = 30 cells in each group). (F) Western blotting analysis of p-DRP1(S616), DRP1, and Tom 20 proteins in HA-VSMCs and HC-VSMCs treated with DMSO, Oligo, HG, and ox-LDL (n = 3 times each group). (G) Representative image of lysosomes stained with dextran in HA-VSMCs and HC-VSMCs treated with DMSO (Ctrl.), bafilomycin A1 (BafA1, 1 μM), HG (20 mM), and ox-LDL (50 μg/mL) for 24 h. (H) Quantitative analysis of the changes in the number and fluorescence intensity of lysosomes in HA-VSMCs and HC-VSMCs treated with DMSO, BafA1, HG, and ox-LDL (n = 30 cells in each group). (I) Western blotting analysis of lysosomal-associated membrane protein 1 (LAMP1), light chain 3 (LC3)-1, and LC3-II in HA-VSMCs and HC-VSMCs treated with DMSO, BafA1, HG, and ox-LDL (n = 3 times each group). *p < 0.05, **p < 0.01, and ***p < 0.001. Scale bar = 10 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)