apolipoprotein e knockout apoe ko mice  (Charles River Laboratories)

Journal: Cells

Article Title: Human FGF1 ΔHBS Gene Therapy as Treatment for Metabolic Dysfunction-Associated Steatohepatitis in ApoE-KO Mice

doi: 10.3390/cells15050387

Figure Lengend Snippet: Induction of a MASH mouse model and experimental design for AAV8-hFGF1 ΔHBS intervention. ApoE-KO mice were fed a HFHC diet for 4 weeks and then administered AAV8-hFGF1 ΔHBS at doses of 4 × 10 9 or 2 × 10 10 vg/mouse. Chow-fed control mice received 2 × 10 10 vg of AAV8-Control. The mice received additional 10 weeks HFHC diet or chow. ( A ) Schematic illustration of the experimental timeline, including HFHC diet feeding, virus injection, IPGTT, IPITT and DEXA scan. After euthanasia, liver tissues were collected and ( B , C ) relative mRNA of hFGF1 ΔHBS and mFGF1 were detected by RT-qPCR, with 18s rRNA used as housekeeping gene; ( D – F ) protein expression of hFGF1 ΔHBS , mFGF1, ZsGreen, and Flag were detected by Western blot, with β-actin used as the loading control, and quantified by densitometry. FGF1-KO mice were used as a negative control to identify mFGF1 band, and different exposure (Ex.) times were applied to distinguish hFGF1 ΔHBS and mFGF1 bands. Data are presented as means ± SEM (n = 3–7). Statistical significance was defined as p < 0.05.

Article Snippet: Ten-week-old male ApoE-KO mice purchased from The Jackson Laboratory (Bar Harbor, ME, USA) were used to establish the MASH mouse model.

Techniques: Control, Virus, Injection, Quantitative RT-PCR, Expressing, Western Blot, Negative Control

Journal: Cells

Article Title: Human FGF1 ΔHBS Gene Therapy as Treatment for Metabolic Dysfunction-Associated Steatohepatitis in ApoE-KO Mice

doi: 10.3390/cells15050387

Figure Lengend Snippet: AAV8-hFGF1 ΔHBS treatment does not significantly alter systemic metabolic parameters. ApoE-KO mice were fed and treated as described in . ( A ) Body weight and ( B ) blood glucose levels were monitored over time. During the final two weeks of the study, ( C ) intraperitoneal glucose tolerance test (IPGTT) and ( D ) area under the curve (AUC) of blood glucose during IPGTT, as well as ( E ) intraperitoneal insulin tolerance test (IPITT) and ( F ) area under the curve (AUC) of blood glucose during IPITT, were assessed. Body composition was analyzed by dual-energy X-ray absorptiometry (DEXA) scan, including ( G ) lean mass (%), ( H ) lean mass (g), ( I ) fat mass (%), and ( J ) fat mass (g). After euthanasia, plasma was collected for measurement of ( K ) triglycerides and ( L ) total cholesterol. Data are presented as means ± SEM (n = 6–7). Statistical significance was defined as p < 0.05. * p < 0.05 for Chow AAV8-Control vs. HFHC AAV8-Control.

Article Snippet: Ten-week-old male ApoE-KO mice purchased from The Jackson Laboratory (Bar Harbor, ME, USA) were used to establish the MASH mouse model.

Techniques: Clinical Proteomics, Control

Journal: Cells

Article Title: Human FGF1 ΔHBS Gene Therapy as Treatment for Metabolic Dysfunction-Associated Steatohepatitis in ApoE-KO Mice

doi: 10.3390/cells15050387

Figure Lengend Snippet: AAV8-hFGF1 ΔHBS treatment attenuates lipid accumulation. ApoE-KO mice were fed and treated as described in . Following euthanasia, ( A ) liver size, ( B ) liver weight, and ( C ) the percentage of liver weight to body weight were measured. Liver sections were subjected to ( D ) hematoxylin–eosin (H&E, upper panel) staining and Oil Red O (red color, middle and lower panel) staining. Hepatic contents of ( E ) triglycerides and ( F ) total cholesterol were quantified. Plasma markers of liver injury, ( G ) ALT and ( H ) AST, were measured. Data are presented as means ± SEM (n = 6–7). Statistical significance was defined as p < 0.05.

Article Snippet: Ten-week-old male ApoE-KO mice purchased from The Jackson Laboratory (Bar Harbor, ME, USA) were used to establish the MASH mouse model.

Techniques: Staining, Clinical Proteomics

Journal: Cells

Article Title: Human FGF1 ΔHBS Gene Therapy as Treatment for Metabolic Dysfunction-Associated Steatohepatitis in ApoE-KO Mice

doi: 10.3390/cells15050387

Figure Lengend Snippet: AAV8-hFGF1 ΔHBS treatment attenuates hepatic inflammation. ApoE-KO mice were fed and treated as described in . Liver sections were analyzed by ( A – D ) immunohistochemical staining to assess macrophage (CD68 + , arrowhead) and neutrophil (Ly6G + , arrowhead) infiltration and TNFα expression (arrowhead), along with corresponding quantification. ( E – I ) Relative mRNA levels of TNFα, IL-1β, CXCL10, CCL2 and ICAM were measured by RT-qPCR, with 18s rRNA used as the housekeeping gene. Data are presented as means ± SEM (n = 6–7). Statistical significance was defined as p < 0.05.

Article Snippet: Ten-week-old male ApoE-KO mice purchased from The Jackson Laboratory (Bar Harbor, ME, USA) were used to establish the MASH mouse model.

Techniques: Immunohistochemical staining, Staining, Expressing, Quantitative RT-PCR

Journal: Cells

Article Title: Human FGF1 ΔHBS Gene Therapy as Treatment for Metabolic Dysfunction-Associated Steatohepatitis in ApoE-KO Mice

doi: 10.3390/cells15050387

Figure Lengend Snippet: AAV8-hFGF1 ΔHBS treatment attenuates fibrosis. ApoE-KO mice were fed and treated as described in . Liver sections were analyzed by ( A – D ) Sirius Red (red color, upper panel) and Masson’s trichrome (blue color, middle panel) staining to evaluate collagen deposition, and immunohistochemical staining to assess Col1a1 protein expression (brown color, lower panel), along with corresponding quantification. ( E – J ) Relative mRNA levels of TGFβ, CTGF, Col1a1, Col1a2, Col13a1 and Desmin were detected by RT-qPCR, 18s rRNA used as the housekeeping gene. Data are presented as means ± SEM (n = 6–7). Statistical significance was defined as p < 0.05.

Article Snippet: Ten-week-old male ApoE-KO mice purchased from The Jackson Laboratory (Bar Harbor, ME, USA) were used to establish the MASH mouse model.

Techniques: Staining, Immunohistochemical staining, Expressing, Quantitative RT-PCR

Journal: Cells

Article Title: Human FGF1 ΔHBS Gene Therapy as Treatment for Metabolic Dysfunction-Associated Steatohepatitis in ApoE-KO Mice

doi: 10.3390/cells15050387

Figure Lengend Snippet: AAV8-hFGF1 ΔHBS treatment improves dysregulated lipid synthesis and transport. ApoE-KO mice were fed and treated as described in . ( A – C ) Relative mRNA levels of SCD1, CPT1α and CD36 were detected by RT-qPCR, 18s rRNA used as the housekeeping gene. ( D – G ) Protein expression levels of SCD1, CPT1α and CD36 were assessed by Western blot, with β-actin as the loading control, and quantified by densitometric analysis. Data are presented as means ± SEM (n = 5). Statistical significance was defined as p < 0.05.

Article Snippet: Ten-week-old male ApoE-KO mice purchased from The Jackson Laboratory (Bar Harbor, ME, USA) were used to establish the MASH mouse model.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Control

Journal: Cells

Article Title: Human FGF1 ΔHBS Gene Therapy as Treatment for Metabolic Dysfunction-Associated Steatohepatitis in ApoE-KO Mice

doi: 10.3390/cells15050387

Figure Lengend Snippet: AAV8-hFGF1 ΔHBS treatment exerts no significant effects on hepatic proliferation. ApoE-KO mice were fed and treated as described in . Liver sections were analyzed by ( A ) immunohistochemical staining for PCNA and Ki67 (arrowhead), along with ( B , C ) corresponding quantification. Data are presented as means ± SEM (n = 6–7). Statistical significance was defined as p < 0.05.

Article Snippet: Ten-week-old male ApoE-KO mice purchased from The Jackson Laboratory (Bar Harbor, ME, USA) were used to establish the MASH mouse model.

Techniques: Immunohistochemical staining, Staining