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. 2022 Jul 1;14(4):925–944. doi: 10.1016/j.jcmgh.2022.06.008

Figure 3.

Figure 3

Induction of lysosomal membrane permeabilization/rupture in hepatocytes by HNE.A, Changes in lysosomes of HepG2 cells after the addition of EPI and HNE were observed by time-lapse imaging using LysoTracker. Blue, Hoechst; red, LysoTracker. Square on the top right indicates morphological changes that were observed on bright field imaging. B, Electron microscopy images of HepG2 cells. Black squares show magnified images of membrane-bound lysosomes in the control (Cont) and disruption of the lysosomal limiting membrane in HNE-treated HepG2 cells (HNE), respectively. L, lysosome. C, HepG2 cells were treated with 25 μM HNE, and imaged 0, 2, and 6 hours later. Blue, DAPI; green, CTSB; red, LysoTracker; yellow, merge. D, Area occupied by LysoTracker and CTSB for each particle in HepG2 cells at a 20× image. The analysis was performed with 10 images in each group. E, Images show immunoreactivity of HepG2 cells for CTSB and LAMP2 before (0) and 6 hours after the treatment of 25 μM HNE. Blue, DAPI; green, CTSB; red, LAMP2; yellow, merge. F, Electron microscopy images of Huh-7. Black squares show magnified images of membrane-bound lysosomes in the control (Cont) and disruption of the lysosomal limiting membrane in HNE-treated Huh7 cells (HNE), respectively. L, lysosome. G, Huh-7 cells were treated with 25 μM HNE, and imaged 0, 2, and 6 hours later. Blue, DAPI; green, CTSB; red, LysoTracker; yellow, merge. H, Area occupied by LysoTracker and CTSB for each particle in Huh7 cells at a 20× image of Huh7. I, Images show immunoreactivity of Huh-7 cells for CTSB and LAMP2 before (0) and 6 hours after the treatment of 25 μM HNE. Blue, DAPI; green, CTSB; red, LAMP2; yellow, merge.