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. 2021 Aug 17;48(4):190. doi: 10.3892/ijmm.2021.5023

Figure 4.

Figure 4

N-3 PUFAs inhibit IL-11 production by downregulating ERK1/2 pathway. (A-C) Hepatocytes were isolated from WT and fat-1 transgenic mice (n=5) at the indicated time after 400 mg/kg APAP administration. (A) The mRNA expression of Fra-1 was evaluated by reverse transcription-quantitative PCR and expressed as a ratio to GAPDH. (B) The protein level of Fra-1 was determined by western blotting analysis. (C) The protein levels of pERK, ERK and GAPDH were detected by western blotting analysis. (D-F) Primary hepatocytes were isolated from WT and fat-1 mice (n=5) and stimulated with 20 mM APAP for 24 h in vitro. In some cases, the cells were pretreated with 50 µM U0126 for 2 h before APAP stimulation. (D) Cellular apoptosis was measured by Annexin V-PI staining. (E) The culture supernatants were collected for evaluating IL-11 production through ELISA. (F) The protein level of Fra-1 in hepatocytes was confirmed by immunoblotting analysis. **P<0.01. NS, not significant; N-3 PUFAs, omega-3 polyunsaturated fatty acids; APAP, acetaminophen; WT, wild-type; Fra-1, Fos-like-1; p, phosphorylated.