Table 1.
List of primer sequences, 5'-3'.
Fig 1.
Male mice consume higher amounts of alcohol.
Male and female mice were subjected to the NIAAA model. (A) Average daily consumption of the 5% Lieber-DeCarli diet per mouse. (B) Serum blood alcohol content (BAC) at the time of sacrifice. * p<0.05, ** p<0.01, *** p<0.001. Data are represented as mean ± SEM. EtOH-fed: Alcohol-fed.
Fig 2.
Female mice have more severe liver injury than male mice.
(A) Liver-to-body weight ratio. (B) Serum alanine aminotransferase (ALT) levels at the time of sacrifice. (C) Liver triglyceride content. (D) Representative Oil Red O staining of liver sections. Scale bar is set to 100 μm and magnification is at 100x. Quantitation was done with Fiji. (E) Liver mRNA levels of TNFα, IL-6, CCL2, F4/80, and Ly6C were quantified by qPCR. * p<0.05, ** p<0.01, *** p<0.001. Data are represented as mean ± SEM.
Fig 3.
Female mice have a higher degree of adipose tissue inflammation.
Periogonadal adipose tissue mRNA levels of (A) TNFα, IL-6, and CCL2 and (B) F4/80, CD68, CD11b, CD11c, and CCR2 were quantified by qPCR. (C) H&E staining of adipose tissue. Scale bar is set to 100 μm and magnification is at 100x. (D) Serum adiponectin levels were measured by ELISA.* p<0.05, ** p<0.01, *** p<0.001. Data are represented as mean ± SEM.
Fig 4.
Increased adipose tissue inflammation is due to innate immune cells.
(A). BMDMs were cultured in the presence of 25mM ethanol for 5 days and then stimulated with 100ng/mL LPS for 18 hours. TNFα was measured in the cell supernatants by ELISA. (B) Serum endotoxin levels at time of sacrifice. (C) Adipose mRNA levels of TLR2, TLR3, TLR4, TLR9, and MyD88 were quantified by qPCR. * p<0.05, ** p<0.01, *** p<0.001. Data are represented as mean ± SEM.