FIG 4.

Kaempferol suppresses the MyD88-dependent inflammatory response downstream of TLR2/4. (A) Western blot analysis of NF-κB and MAPK signaling cascades in MPMs stimulated with TLR4 ligand LPS (1 mg/mL) in the presence of DMSO or 32 mg/mL kaempferol. (B) The levels of P-p65, P-IKK, P-p38 and P-JNK relative to β-actin determined using densitometry are shown. (C) The levels of cytokines TNF-α, IL-1β, and IL-6 in the culture supernatants were detected using ELISA. (D) The transcriptions of these proinflammatory genes were determined by RT-qPCR. Notably, all the genes were normalized to the housekeeping gene GAPDH. (E) Western blot analysis of NF-κB and MAPK signaling cascades in MPMs stimulated with the TLR2 ligand Pam3CSK4 (1 mg/mL) in the presence of DMSO or 32 mg/mL kaempferol. (F) The levels of P-p65, P-IKK, P-p38 and P-JNK relative to β-actin determined using densitometry are shown. (G) The levels of cytokines TNF-α, IL-1β, and IL-6 in the culture supernatants were detected using ELISA. (H) The transcriptions of these proinflammatory genes were determined by RT-qPCR. Notably, all thegenes were normalized to the housekeeping gene GAPDH. Similar results were obtained from three independent experiments. Data are presented as means 6 SEM (n = 3). **, P, 0.01 compared to the vehicle-treated group.