Journal: iScience

Article Title: Aquaporin-8 ameliorates hepatic steatosis through farnesoid X receptor in obese mice

doi: 10.1016/j.isci.2023.106561

Figure Lengend Snippet: Downregulation of AQP8 in the livers of NAFLD patients and obese mice and effects of hepatic AQP8 overexpression in vitro Hepatic mRNA and protein levels of AQP8 decreased in patients and obese mouse model: (A) NAFLD patients (n = 8 per group), (B) db/db mice (n = 6 per group), (C) high fat diet (HFD) mice (n = 7 per group), (D) short-term HFD mice (n = 5 per group) (See also Figure S2 A), (E) HepG2 cells without or with FFA(OA:PA = 2:1) treatment for the indicated time(See also Figure S2 B). Primary mouse hepatocytes (PMHs) and HepG2 cells were transfected with either ad-GFP or ad-AQP8 with or without FFA treatment. (F) AQP8 mRNA expression of PMHs. (G) Analysis of AQP8 and lipogenic genes expression (See also Figure S2 C). (H) Oil red O staining (200 X) of HepG2 cells. (I) TG measurement of PMHs. The data represent the mean ± SEM values. ∗p < 0.05 ∗∗p < 0.01 ∗∗∗p < 0.001.

Article Snippet: Mouse primary hepatocytes and HepG2 cells (AmericanType Culture Collection, China) were cultured in DMEM supplemented with 10% of fetal bovine serum, 1% of L-glutamine, and 1% of penicillin-streptomycin solution and maintained in a humidified atmosphere of 5% CO2 at 37°C.

Techniques: Over Expression, In Vitro, Transfection, Expressing, Staining

Journal: iScience

Article Title: Aquaporin-8 ameliorates hepatic steatosis through farnesoid X receptor in obese mice

doi: 10.1016/j.isci.2023.106561

Figure Lengend Snippet: FXR regulates AQP8 expression and FXR knockout inhibits the improvement of AQP8 in NAFLD (A) PMHs were treated with DMSO, PPARα agonist Fenofibrate, PPARβ/δ agonist GW0742, PPARγ agonist Rosiglitazone, FXR agonist GW4064, LXR agonist T0901317 and RT–PCR analysis of AQP8 gene expression. (B) Dual-luciferase reporter assay was conducted to determine the effect of FXR agonist GW4064 on AQP8 activity. (C) Dual-luciferase reporter assay was conducted to determine the effect of FXR overexpression on AQP8 activity. (D) The interaction of FXR with AQP8 promoter containing putative FXRE was confirmed by chromatin immunoprecipitation (ChIP) in HepG2 cells (n = 3 individual experiments) using an FXR antibody, IgG was used as a negative control. (E) Relative AQP8 mRNA levels in the livers of FXR wild-type (WT) and knockout (KO) mice. (F) FXR-WT and FXR-KO were fed an HFD for two weeks and then i.v injected with either ad-GFP or ad-AQP8 and relative AQP8 mRNA levels in the livers of three groups. (G–J) Serum levels of TBA, TG, TC, LDL-c. (K) The analysis of liver weight in three groups. The count is liver weight per kilogram of body weight. (L) Quantification of hepatic TG contents. (M) Histological analysis of hematoxylin-eosin-stained liver sections, magnification: 200×, scale bar, 100 μm. The data represent the mean ± SEM values. ∗p<0.05 ∗∗p<0.01 ∗∗∗p<0.001.

Article Snippet: Mouse primary hepatocytes and HepG2 cells (AmericanType Culture Collection, China) were cultured in DMEM supplemented with 10% of fetal bovine serum, 1% of L-glutamine, and 1% of penicillin-streptomycin solution and maintained in a humidified atmosphere of 5% CO2 at 37°C.

Techniques: Expressing, Knock-Out, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Luciferase, Reporter Assay, Activity Assay, Over Expression, Chromatin Immunoprecipitation, Negative Control, Injection, Staining

Journal: iScience

Article Title: Aquaporin-8 ameliorates hepatic steatosis through farnesoid X receptor in obese mice

doi: 10.1016/j.isci.2023.106561

Figure Lengend Snippet:

Article Snippet: Mouse primary hepatocytes and HepG2 cells (AmericanType Culture Collection, China) were cultured in DMEM supplemented with 10% of fetal bovine serum, 1% of L-glutamine, and 1% of penicillin-streptomycin solution and maintained in a humidified atmosphere of 5% CO2 at 37°C.

Techniques: Virus, Software