Norleual inhibits HGF-dependent Met phosphorylation and Gab1 association and
phosphorylation in vitro. a, HEK293 cells were treated for 10 min with HGF
at the indicated concentrations. Lysates were IP with anti-Met (DO-24) and
immunoblotted (IB) with anti-Met or anti-phosphotyrosine (06-427) b, HEK293
cells were treated for 10 min with HGF and/or norleual at the indicated
concentrations. Lysates were IP with anti-Met (DO-24) and IB with anti-Met
(DQ-13) or anti-phosphotyrosine (HAM 1676, lot JLB03). c, HEK293 lysates
were IP with anti-Met (DO-24) and IB with anti-Gab1. d, HEK293 lysates were
IP with anti-Gab1 and IB with anti-Gab1 or anti-phosphotyrosine (HAM 1676).
e, relative amounts (normalized to percentage of total Met in IP) of
quantified IP/IB band intensities for control (black bars), 110 pM HGF
(checkered bars), 110 pM HGF plus 20 pM norleual (white bars), and 20 pM
norleual (gray bars)-treated HEK293 cells for phospho-Met
(n = 4), phospho-Gab1 (n = 4), and Met
associated Gab1 (n = 3). Error bars are standard deviation,
and p values were determined by one-way analysis of
variance followed by Tukey-Kramer multiple comparison test.