VacA-induced Akt phosphorylation leading to GSK3β phosphorylation
at Ser9. A, AZ-521 cells were incubated with 120 nm
VacA or heat-inactivated VacA (iVacA) at 37 °C for the indicated
times. After incubation, cell lysates were analyzed by SDS-PAGE (10% gels),
followed by Western blot analyses using anti-phospho-Thr-308 Akt,
phospho-Ser-473 Akt, and Akt antibodies. Results are representative of three
independent experiments. Quantification of phospho-Thr-308 Akt and
phospho-Ser-473 Akt obtained with VacA (filled bars) and iVacA
(open bars) was performed by densitometry. Data are means ±
S.E. of values from triplicate experiments, with an n = 3 per
experiment. Statistical significance: *, p < 0.01.
B, AZ-521 cells were incubated with 120 nm VacA
(filled bars) or iVacA (open bars) at 37 °C for
indicated times. Cell lysates were prepared at indicated incubation times and
subjected to Western blot analyses using anti-phospho-Ser-9 GSK3β and
GSK3β antibodies. Results are representative of three independent
experiments. Quantification of phospho-Ser-9 GSK3β obtained with VacA
(filled bars) and iVacA (open bars) was determined by
densitometry. Data are means ± S.E. of values from triplicate
experiments, with an n = 3 per experiment. Statistical significance:
*, p < 0.05.