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

Figure 7.

Figure 7

Administration of Alda-1 in CDAA mice suppresses liver fibrosis.A, Hematoxylin and eosin staining and HNE immunostaining of liver tissue from the control mice fed the standard diet (Cont), CDAA mice (CDAA) and Alda-1-treated CDAA mice (CDAA+Alda-1) for 8 weeks. B, The expressions of liver HNE protein adducts in CDAA mice (CDAA) and CDAA mice with Alda-1 treatment (CDAA+Alda-1) for 4 (4w) and 8 (8w) weeks were evaluated by a Western blotting analysis. C, Bands of panel B are quantified and shown as relative fold ratios. D, Comparison of histopathological scores of CDAA mice (CDAA) and Alda-1 treated CDAA mice (CDAA+Alda-1) for 8 weeks, such as steatosis score, lobular inflammation score (Lobular inf), ballooning score, and NAFLD activity score (NAS) is shown. E, Sirius red staining of liver tissue from CDAA mice (CDAA) and Alda-1-treated CDAA mice (CDAA+Alda-1) for 8 weeks. F, Area occupied by Sirius red staining in a 20× image were compared between the CDAA mice (CDAA) and Alda-1 treated CDAA mice (CDAA+Alda-1) for 8 weeks. The area was calculated for each of the 10 images and averaged. G, Real-time PCR analysis of IL-1β, IL-6, tumor necrosis factor (TNF), and toll-like receptor (TRL)-4 expression in CDAA mice (blue circle) and Alda-1 treated CDAA mice (red triangle) for 4 and 8 weeks. H, Immunofluorescence staining of livers in control mice (Cont) and the CDAA mice (CDAA). Blue, DAPI; green, CTSB; red, LAMP2. I, Electron microscopy images of liver tissue collected from control mice (Cont) and CDAA mice (CDAA) for 8 weeks. The black arrow shows the disruption of the lysosomal limiting membrane. L, lysosome. J, Immunofluorescence LAMP2 staining of liver tissue from CDAA mice (CDAA) and Alda-1-treated CDAA mice (CDAA+Alda-1) for 8 weeks. K, The area of particles stained with LAMP2 in CDAA mice (CDAA) and Alda-1 treated CDAA mice (CDAA+Alda-1) for 4 weeks is shown. L, The number of stained areas that were 10 μm2 or larger in panel K is shown. M, The area of particles stained with LAMP2 in CDAA mice (CDAA) and Alda-1 treated CDAA mice (CDAA+Alda-1) for 8 weeks is shown. N, The number of stained areas that were 10 μm2 or larger in panel M is shown.