Fig. 4. Pim1 promotes the phosphorylation and nuclear translocation of IRF3.

a Ability of Pim1 to affect LPS-induced phosphorylation of TLR4 upstream kinases or downstream IRF3. WT or Pim1^−/− BMDMs were treated with LPS (100 ng ml⁻¹) for the indicated times and were then subjected to immunoblot analysis of phosphorylated TBK1, IRF3, STAT1, NF-κB p65, ERK, p38 and JNK. b, c Ability of Pim1 to promote the nuclear translocation of IRF3. WT and Pim1^−/− BMDMs were treated with LPS (100 ng ml⁻¹) for 1 h. The cytoplasmic and nuclear fractions of these cells were isolated and subjected to immunoblot analysis b, or the intact cells were used for immunofluorescence analysis of endogenous IRF3 with DAPI counterstaining of nuclei. Scale bars, 10 μm. All data are from at least two independent experiments with similar results.