Figure 5 | Scientific Reports

Figure 5

From: Micheliolide inhibits LPS-induced inflammatory response and protects mice from LPS challenge

Figure 5

MCL inhibits the activation of NF-κB signaling pathway after LPS stimulation.

Raw264.7 cells (1 × 106/well) was plated in 6-well plates overnight and stimulated with LPS (100 ng/mL) with or without MCL (10 μM) for the indicated time periods. (a) Phospho-ERK, p-JNK, p-p38 and the corresponding total ERK, JNK and p38 were detected by Western blot. Similar results were obtained in three independent experiments. (b) Phospho-IκBα, total IκBα and β-Actin were detected in Raw264.7 cells by Western blot. Similar results were obtained in three independent experiments. (c) Human monocytic cell line THP-1 (1 × 106/well) was plated in 6-well plates overnight with PMA (10 ng/mL) and stimulated with LPS and MCL as indicated. Phospho-IκBα, total IκBα and β-Actin were detected by Western blot. Similar results were obtained in three independent experiments. (d) Raw264.7 cell line was cotransfected with NF-κB luciferase reporter plasmid and pRL-TK-Renilla-luciferase plasmid. After 30 h, cells were stimulated with LPS (100 ng/mL) for 24 h and luciferase activities were measured. The NF-κB luciferase activities were presented as fold increase.

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