FIGURE 2.
Kinetics of IL-23-induced STAT3 activation. A, Ba/F3-gp130 cells stably transduced with cDNAs for murine IL-23R and murine IL-12Rβ1 were washed three times with PBS and starved for 2 h in serum-free DMEM. 1 × 106 cells were stimulated for the times indicated with 0.2% HIL-6 or HIL-23, harvested by centrifugation, fixed in 2% (w/v) paraformaldehyde, and permeabilized in 90% (v/v) methanol. Cells were stained for phospho-STAT3 and STAT3 overnight and analyzed by flow cytometry. B, in parallel, cellular lysates of the respective Ba/F3-gp130 cells, stimulated for the times indicated with 0.2% HIL-6 or HIL-23, were prepared and analyzed by Western blot using antibodies specific for phospho-STAT3 and STAT3.