Fig. 3

EEST inhibits protein expression of c-Fos and both mRNA and protein expression of NFATc1. a BMMs were pretreated with or without EEST (50 μg/mL) for 1 h and then stimulated with M-CSF (30 ng/mL) and RANKL (50 ng/mL) for the indicated times. The cell lysates were analyzed by western blotting with antibodies against c-Fos, NFATc1, and GAPDH. b BMMs were stimulated with RANKL (50 ng/mL) and M-CSF (30 ng/mL) in the presence or absence of EEST (50 μg/mL) for the indicated times. Total RNA was isolated from cells using the QIAzol reagent, and mRNA expression of c-Fos and NFATc1 was determined using quantitative real-time RT-PCR. ***P < 0.001 vs. control in the indicated time