Fig. 3

WEMV downregulates c-Fos and NFATc1 expression. BMMs were pretreated with WEMV (160 μg/mL) for 3 h and cultured in differentiation medium for the indicated times as described in the Methods. Total RNA or protein was obtained at the indicated time points. a Total cell lysates (30 μg) were separated by SDS-PAGE electrophoresis and transferred to PVDF membranes. Western blot analysis was performed with antibodies specific for c-Fos (top), NFATc1 (middle), and β-actin (bottom). b qRT-PCR analysis of c-Fos, NFATc1, MafB, and Id2 was performed at the indicated time points. Each mRNA level was normalized to that of HPRT. c BMMs were infected with retrovirus vector harboring GFP (control) or CA-NFATc1 in the presence of polybrene (6 μg/mL) for 8 h. The infected cells were cultured in α-MEM medium containing puromycin (2 μg/mL) for 5 days and then cultured in differentiation medium containing WEMV (160 μg/mL) for 4 days. TRAP-positive stained MNCs were counted after TRAP staining under an inverted microscope (40× magnification). **p < 0.01 versus control. NS; not significance