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Fig. 7 | Journal of Experimental & Clinical Cancer Research

Fig. 7

From: Alcoholic fatty liver disease inhibited the co-expression of Fmo5 and PPARα to activate the NF-κB signaling pathway, thereby reducing liver injury via inducing gut microbiota disturbance

Fig. 7

Fmo5 and PPARα co-expression alleviates AFLD-induced apoptosis, inflammatory response and the nuclear translocation of NF-κB p65 via inhibiting the activation of NF-κB signaling pathway in vitro. Western blot assay was used to detect the phosphorylation level of NF-κB p65 (a) in vector expressing PPARα and Fmo5 transfected L02 cells, b in siRNA targeting PPARα and Fmo5 transfected L02 cells, and (c) in siRNA targeting PPARα and vector expressing Fmo5. ELISA assay was used to detect TG level (d), AST level (e), ALT level (f), and IL-6/− 4 and TNF-α levels (g) in L02 cells among six groups. h CCK-8 assay was used to detect the cell viability in L02 cells among six groups. Flow cytometry assay was used to detect ROS level (i) and cell apoptosis (j) in L02 cells among the six groups. k Immunofluorescence staining was used to observe the nuclear translocation of NF-κB p65 in L02 cells among six groups. l and m Western blot assay was used to detect the protein levels of Fmo5, PPARα, phospho-NF-κB p65 and NF-κB p65 in CHS-828 treated L02 cells. After CHS-828 pre-treated with L02 cells with AFLD for 4 h, siRNA-Fmo5 and siRNA-PPARα were co-transfected into cells, the level of phospho-NF-κB p65 (n) was detected by Western blot assay, cell viability (o) was detected by CCK-8 assay, the levels of TG, AST, ALT (p), IL-6, IL-4, and TNF-α level (q) were detected by ELISA assay, cell apoptosis (r), and ROS level (s) were detected by Flow cytometry assay. Data are presented as the mean ± standard deviation. a P < 0.01 vs. NC/or siNC+NC/or Con group, b P < 0.01 vs. PPARα/or Fmo5/or siPPARα/or AFLD group, c P < 0.01 vs. Fmo5/or siFmo5/or siPPARα/or AFLD+siFmo5 group, d P < 0.01 vs. AFLD+siFmo5+ PPARα group, and e P < 0.01 vs. AFLD+Fmo5+ siPPARα group

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