Fig 1: eIF2α-mediated integrated stress responses verified in carotid stable and vulnerable plaques. (A) Representative specimens of stable and vulnerable plaques stained with H&E, and immunohistochemistry (IHC) of VSMCs with organelle markers, including α-SMA, PERK, XBP1, GRP78, cytochrome C (Cyt C), LAMP1, and eIF2α. (B) Quantitative analysis of the expression levels of α-SMA, PERK, XBP1, GRP78, Cyt C, LAMP1, and eIF2α (n = 15 cases in each group). *p < 0.05, **p < 0.01, and ***p < 0.001. Scale bar = 200 μm, 1 cm, 2 cm, respectively.
Fig 2: Eukaryotic translation initiation factor (eIF)-2α integrates the stress responses of multiple organelles to regulate atherosclerotic plaque progression. (A) Representative image of ER labeled with GFP-Sec61β and mitochondria stained with deep red Mitotracker in HC-VSMCs treated with DMSO (Ctrl.), thapsigargin (Tg; 5 μM), Oligo (1 μM), and combined Tg and oligo for 24 h. (B) Quantitative analysis of the percentage of cells with ER whorls and the aspect ratio in HC-VSMCs treated with DMSO (Ctrl.), Tg, Oligo, and combined Tg and Oligo (n = 30 cells per group). (C) Representative image of ER labeled with GFP-Sec61β and mitochondria stained with deep red Mitotracker in HC-VSMCs treated with DMSO (Ctrl.), Tg (5 μM), ISRIB (100 nM), Oligo (1 μM), and their combination for 24 h. (D) Quantitative analysis of the percentage of cells with ER whorls and the aspect ratio in HC-VSMCs treated with DMSO, DTT, ISRIB, Oligo, and their combination (n = 30 cells per group). (E) Western blotting analysis of GRP78, XBP1-u, XBP1-s, p-DRP1(S616), LC3-I, and eIF2α in HC-VSMCs treated with DMSO (Cntrl.), Tg, Oligo, and combined Tg and Oligo (n = 3 in each group). (F) Western blotting analysis of GRP78, XBP1-u, XBP1-s, p-DRP1(S616), LC3-I, and eIF2α in HC-VSMCs treated with DMSO, Tg, ISRIB, Oligo, and their combination (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.)
Supplier Page from Sino Biological, Inc. for Human HSPA5 Gene ORF cDNA clone expression plasmid