Figure 6-6923.

The effects of EYFP-p62WT and EFYP-p62ΔUBA overexpression on RANKL-induced phosphorylation of ERK in RAW stable cell lines. RAW264.7 cells stably transfected with pcDNA3.1, EYFP-p62WT, and EFYP-p62ΔUBA were stimulated with 100 ng/ml of RANKL for the indicated time. On stimulation, whole cell extracts were analyzed for phosphorylation of ERK 1 and 2 proteins by Western blotting with monoclonal anti-p-ERK antibody at 1/1000 dilution, followed by enhanced chemiluminescence. The same membrane was stripped and reprobed with an antibody for β-tubulin, which served as an internal control for differences in loading and transfer. A: The levels of ERK 1/2, and β-tubulin proteins are shown. B and C: Levels of phosphorylation of ERK proteins are shown as the ratio of phosphorylated ERK to β-tubulin. Each bar is the mean ± SEM from three independent experiments (***P < 0.001).