Figure 8

Macrophage migration inhibitory factor induces CD74-dependent activation of the AMPK-FOXO3a signaling pathway. (A, B) Representative images of western blots of AMP-activated protein kinase (AMPK) and phospho-AMPK in mesenchymal stem cells (MSCs) transfected with CD74-small interfering RNA (siRNA) or scrambled small interfering RNA (siRNA-NT) before pretreatment with macrophage migration inhibitory factor (MIF) (100 ng/ml) and incubated in normal conditions for the indicated time. Fold-changes were calculated normalized to AMPK. Each column represents mean ± standard deviation from three independent experiments; *P <0.05 versus control; ^△ P <0.05 versus siRNA-CD74. (C, D) Representative images of western blots of AMPK and β-actin in MSCs transfected with siRNA against AMPK genes, and siRNA-NT. Each column represents mean ± standard deviation from three independent experiments; *P <0.05 versus siRNA-AMPK. Fold-changes were calculated normalized to β-actin. (E, F) Representative images of western blots of Forkhead box class O 3a (FOXO3a) and phospho-FOXO3a in MSCs transfected with siRNA-AMPK or siRNA-NT before pretreatment with MIF (100 ng/ml) and incubated under normal conditions for the indicated time. Fold-changes were calculated normalized to FOXO3a. Each column represents mean ± standard deviation from three independent experiments; *P <0.05 versus control; ^△ P <0.05 versus siRNA-AMPK. (G, H) Representative images of western blots of FOXO3a and β-actin in MSCs transfected with siRNA against FOXO3a genes, and siRNA-NT. Fold-changes were calculated normalized to β-actin. Each column represents mean ± standard deviation from three independent experiments; *P <0.05 versus siRNA-FOXO3a.