SFK inhibition decreasing
DDR2 phosphorylation and suppressing
DDR2 mutated cancer cell lines. (A) DDR2 was ectopically expressed
in 293T cells, and phosphorylation was measured by Western blotting
with antiphosphotyrosine (4G10) after immunoprecipitation with an
anti-DDR2 antibody. Cells were treated with the indicated concentration
of 1 in combination with or without 1 μM of saracatinib.
(B) Proliferation of NCI-H2286 and HCC-366 cells grown for 5 days
in the presence of compound 1 in combination with DMSO
or 1 μM of saracatinib or in the presence of different concentrations
of saracatinib with DMSO. Graph shows mean ± SD from a single
experiment representative of three independent experiments with three
replicates per treatment per experiment. (C) DDR2 WT or SRC WT was
ectopically expressed in 293T cells. Cells were treated with saracatinib.
Receptor phosphorylation was measured by Western blotting with antiphosphotyrosine
after immunoprecipitation with an anti-DDR2 antibody. Western blotting
for anti-DDR2, anti-t-SRC, anti-p-SFK, and antiactin was performed
with the same lysate. (D) DDR2 WT, DDR2 GK, SRC WT, or SRC GK was
ectopically expressed in 293T cells. Cells were treated with a vehicle
or dasatinib. Western blotting with antiphosphotyrosine, anti-DDR2,
or total SRC was performed after immunoprecipitation with an anti-DDR2
antibody.