Fig. 6. The activation state of the RhoA-GTPase. (A) Immunoblots of control
pull-downs using the human form of rhotekin-RBD showing that it is specific
for the activated form of RhoA. Approximately 104 SW13 cells were
seeded in 5 cm dishes and left untreated or transfected with the vector as
indicated above the lanes for 24 hours prior to the pull-down. Myc-tagged
mutant forms of RhoA were detected using the anti-myc-9E antibody. (B) The
activation states of endogenous RhoA in SW13 cells, a BRG1-expressing clone
and a BRG1-K798R-expressing clone were measured 24 hours after seeding at
60% confluence. The protein concentration of each sample was measured in
the crude lysates and the protein input adjusted accordingly. The RhoA protein
was detected using anti-RhoA antibodies. (C) The activation state of RhoA in
SW13 cells, BRG1 and BRG1-K798R-expressing clones seeded at 60%
confluence, starved for 18 hours in serum-free medium and stimulated with 5%
serum for 20 minutes as indicated above the lanes is shown. (The experiment
presented is representative of three experiments.)
