Fig. 6

ISO inhibits RANKL-induced activation of MAPK and PI3K-AKT1 signaling cascades and down-regulated the expression of downstream c-Fos and NFATc1. A, B RAW264.7 cells were pretreated with ISO, and then total protein was extracted after stimulation with 50 ng/mL RANKL for 0, 15, 30, and 60 min. Finally, western blot analysis was performed against ERK and p-ERK, JNK and p-JNK, p38 and p-p38, p-PI3K and PI3K, p-AKT1 and AKT1. C RAW264.7 cells were stimulated with RANKL for the indicated times in the presence or absence of ISO at 100 μM respectively, and analyzed by western blot using specific antibodies against c-Fos, NFATc1, and β-actin. D–J Quantitative densitometric analysis of phospho-ERK relative to ERK, phospho-JNK relative to JNK, phospho-p38 relative to p38, phospho-PI3K relative to PI3K, phospho-AKT1 relative to AKT1, and NFATc1 and c-Fos relative to β-actin. K Quantification of relative mRNA expression of specific genes of OCs (NFATc1 and c-Fos). Values were presented as mean ± standard deviation (n = 3); *p < 0.05, **p < 0.01