Figure 2.

Effects of IL-34 on MEK/ERK and JNK/c-Jun signaling pathways in JB6 Cl41 cells. (A–D) Cells were serum starved for 24 h, treated with indicated doses of IL-34 for 30 min (A,C) or 10 ng/mL IL-34 for the indicated times (B,D), harvested, and lysed. The lysates were resolved using SDS-PAGE and immunoblotting analysis was performed using specific antibodies against corresponding proteins. (E) Cells were transfected with mouse siRNA-control and siRNA-CSF1R. At 24 h after transfection, the cells were serum starved for 24 h, treated with 10 ng/mL IL-34 for 15 min or left untreated, harvested, and lysed. Proteins in whole cell lysates were separated using SDS-PAGE and immunoblotted. (F,G) Cells were serum starved for 24 h, pre-treated with different concentrations of PD98059 (F) or SP600125 (G) for 12 h, exposed to 10 ng/mL IL-34 for 15 min, harvested, and lysed. Proteins in whole cell lysates were separated using SDS-PAGE and immunoblotted. (A–F) Blots are representative of an experiment repeated at least three times with the similar result. The numbers below the band represent fold changes in protein levels after normalization to β-actin using densitometric quantification by ImageJ.