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. 2018 Jan 3;6:e4208. doi: 10.7717/peerj.4208

Figure 3. Involvement of SIRT1 in the formononetin-mediated inhibition of LPS-induced HMGB1 release in RAW264.7 cells.

Figure 3

(A) Cells cultured in serum-free medium for 16 h were stimulated with LPS in the presence or absence of formononetin for 24 h. (B) Cells were transfected with siRNA against SIRT1 or control and incubated for 38 h. (C) Cells transfected with or without SIRT1-targeting siRNA for 38 h were exposed to LPS in the presence or absence of formononetin for 24 h. (D, E) Cells pretreated with sirtinol (D) or resveratrol (E) for 30 min were stimulated with LPS in the presence or absence of formononetin for 24 h. Equal volumes of conditioned media or aliquots of whole-cell lysates were analyzed by immunoblotting with the indicated antibodies. Ponceau S staining and β-actin were used as a loading control. Representative blots are provided. The fold changes in the SIRT1/β-actin or HMGB1/Ponceau S ratio relative to that in the untreated group are shown as mean ± SE (n = 3). **p < 0.01 compared with the untreated group; #p < 0.05, ##p < 0.01 compared with the LPS-treated group; †p < 0.05 compared with the LPS plus formononetin-treated group.