3.4. ERK1/2 kinase activation activity of
cp-hFGF7115-114
hFGF7 can trigger Ras-Raf-ERK1/2 MARK pathway signaling by direct
interaction with the FGFR2b receptor, thereby further stimulating
reactions such as epithelial cell proliferation and
migration.40 Based on the previously reported works
for mouse and human cell-based functional analysis of
hFGF7,41, 42 we also evaluated whether the MARK
signaling pathway was activated by recombinant
cp-hFGF7115-114 using a mouse embryonic fibroblast
NIH3T3 cell. NIH3T3 cells were treated with the purified
cp-hFGF7115-114, and ERK1/2 phosphorylation in these
cells was evaluated in a time- and dose-dependent
manner.43 As shown in Figure 4A,
cp-hFGF7115-114 (50 ng/mL) induced clear
phosphorylation of ERK1/2, and its effect decreased in a time-dependent
manner. In addition, cp-hFGF7115-114 also activated
ERK1/2 phosphorylation in a dose-dependent manner that was similar to
the commercially available wild type protein (Figure 4B). These results
provided strong evidence that the CP did not profoundly change the
biological activity of cp-hFGF7115-114 and thus
retained a capability for phosphorylation of ERK1/2 in experimental cell
lines.