Journal: Frontiers in Nutrition
Article Title: High intake of n-6 polyunsaturated fatty acid exacerbates non-alcoholic steatohepatitis by the involvement of multiple metabolic pathways
doi: 10.3389/fnut.2025.1562509
Figure Lengend Snippet: Effects of n-6 PUFA on liver macrophage phenotype in rats with NASH induced by a choline-deficient diet. (A) M1-type Kupffer cells (KCs) identified by double staining: red arrows show CD11c-positive cells, green arrows show CD68-positive cells, and yellow arrows highlight CD11c and CD68 double-positive M1-type KCs (Scale bar – 50 μM). (B) M2-type KCs identified similarly, with red arrows indicating CD163-positive cells, green arrows showing CD68-positive cells, and yellow arrows marking CD163 and CD68 double-positive M2-type KCs (Scale bar – 50 μM). For (A,B) (see ) for full-size photomicrographs. (C) M1/M2 phenotype ratio (unitless), calculated as the proportion of CD68 + CD11c + to CD68 + CD163 + cells. (D) Relative PPAR-γ2 mRNA expression (fold change normalized to GAPDH) in the liver, which is linked to macrophage polarization and inflammation. Data are expressed as mean ±SEM; n = 6/group.
Article Snippet: BCA protein quantitation kit (Boster Bio, Cat # AR0146), RIPA lysis buffer (Boster Bio, Cat # 0105), protease inhibitor cocktails (Boster, Bio Cat # AR1182), phosphatase inhibitor (Boster Bio, Cat # AR1183), color pre-dyed protein marker (Boster Bio, Cat # AR1113), Western-specific primary and secondary antibody diluent (Boster Bio, Cat # AR1017), wash buffer TBS-T (Boster Bio, Cat # AR0195-10), ECL chemiluminescent reagent (Boster Bio, Cat # AR1196), BSA TBS buffer system blocking solution (Boster Bio, Cat # AR0189), NF-κB antibody (Boster Bio, Cat # A01228-1), GAPDH antibody (Boster Bio, Cat # M00227), HRP-conjugated goat anti-rabbit IgG (Boster Bio, Cat # BA1054), CD163 antibody (Boster Bio, Cat # A00812-2), CD11c antibody (Boster Bio, Cat # A00357-3), CD68 antibody (Boster Bio, Cat # BA3638), fluorescent (DyLight 488) labeled goat anti-rabbit IgG (Boster Bio, Cat # BA1127), fluorescent (DyLight 594) labeled goat anti-rabbit IgG (Boster Bio, Cat # BA1142), rat TNF-α ELISA kit (Jiangsu Enzyme Immunoassay Co., Ltd., Cat # MM-0180R1), rat IL-1β ELISA kit (Boster Bio, Cat # EK0393), rat IL-4 ELISA kit (Jiangsu Enzyme Immunoassay Co., Ltd., Cat # MM-0191R1), rat IL-10 ELISA kit (Boster Bio, Cat # EK0418), 25 VD3 assay kit (Roche Diagnostics, Cat # 07028148190), TAOC assay kit (Nanjing Jiancheng, Cat # A015-3-1), MDA assay kit (Nanjing Jiancheng, Cat # A003-1-2), and free fatty acid kit (Kunchuang Biotechnology, Xian, China, Cat # SK125-2).
Techniques: Double Staining, Expressing