Figure 7

STATc deletion delays AdcA dephosphorylation. A. Disruption of adcA has no effect on STATc response in conditions of hyperosmolarity. adcA null cells were treated with 200 mM sorbitol (t = 0). STATc phosphorylation and nuclear translocation were examined by Western blot (7H3 and CP22 antibodies) and immunofluorescence (7H3 antibody) respectively. Scale bar: 10 μm. B and C. STATc disruption leads to a persistence of AdcA phosphorylated status. The STATc knock-out strain (dstC^-), Ax-2 cells (dstC^- parent) and KAx-3 cells overexpressing _mycPTP3 were subjected to 200 mM sorbitol at t = 0 and AdcA phosphorylation was followed by Western blot. The right panels illustrate the level of activation of STATc in these different strains in response to sorbitol (B). The signal intensity of bands 2 + 3 was measured using ImageJ software, expressed as a percentage of the total signal intensity (bands 1 + 2 + 3) and plotted as a function of time. The normalized data are shown for the dstC^- and Ax-2 cells as the mean ± s.e.m. (n = 3) (C). D. Cycloheximide mimics the effect of STATc disruption. KAx-3 cells were treated with 2 mM cycloheximide alone or 30 min prior to treatment with 200 mM sorbitol and AdcA electrophoretic profile was followed by Western blot.