Figure 6

JAK2 mediated an alternative dominant p65 phosphorylation in TGEV infected ST cells. (A) CYT387 diminished JAK2 phosphorylation in the presence and absence of tylophorine in TGEV infected ST cells. Cells were treated with vehicle DMSO, tylophorine, IMD-0354, CYT387, or the combination of tylophorine and CYT387 for 1 h prior to TGEV infection and harvested at 15 m.p.i. for western analyses with the indicated antibodies for JAK1, p-JAK1, JAK2, p-JAK2, JAK3, p-JAK3, Tyk2, p-Tyk2, and GAPDH. The concentrations of IMD-0354 or CYT387 (µM) were used as indicated and 1 µM of for tylophorine was used. (B) JAK2 knockdown diminished the phosphorylation of p65 in TGEV infected ST cells. Control shRNA or shJAK2 were transduced to ST cells to knockdown JAK2 expression. The resultant JAK2-knockdown ST cells or ST cells harbored control shRNA were infected by TGEV respectively at an MOI of 7 in presence or absence of 1 µM tylophorine. Cells were harvested at 5, 10 and 15 m.p.i. of TGEV and the resultant cell lysates were subjected to western analysis with the indicated antibodies for JAK2, p65, p-p65 (S536) and β-actin. Uncropped images for Fig. 6 are presented in Supplementary Figure S11.