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. 2021 Aug 20;42(12):2069–2081. doi: 10.1038/s41401-021-00742-0

Fig. 5. EF activates the AMPK-mediated signaling pathway but fails to deactivate the MAPK pathway.

Fig. 5

RAW264.7 cells (a) and BMDMs (b) were pretreated with EF (0, 3, 10, 30, and 100 μM) for 0.5 h, followed by stimulation with LPS (100 ng/mL) for another 0.5 h. a, b The total protein of cells was extracted, and the expression levels of p-p38, p38, p-JNK, JNK, p-ERK, and ERK were analyzed by Western blot with β-actin as the loading control. RAW264.7 cells (c, e) and BMDMs (d, f) were plated in 6-well plates, pretreated with EF (0, 3, 10, 30, and 100 μM) for 0.5 h, and then challenged with LPS (100 ng/mL) for another 0.5 h. c, d The protein expression levels of p-AMPK and AMPK were measured using Western blot analysis. e, f The ratio of p-AMPK/AMPK was quantified using ImageJ software. g–i RAW264.7 cells pretreated with 5 μM Compd C were treated with EF (100 μM) for 0.5 h and then challenged with LPS (100 ng/mL) for another 24 h. The protein expression levels of iNOS and Nrf2 were measured, β-actin was used as a loading control (g), iNOS/β-actin (h), and Nrf2/β-actin (i) were analyzed by ImageJ software. Data are shown as the mean ± SEM, n = 3, *P < 0.05; **P < 0.01; ***P < 0.001.