FIG 7.

Pim kinases inhibit Tax transactivation activity. (A) 293T cells were transfected with the HTLV-1 LTR luciferase construct along with pcTax and increasing amounts of pcDNA-Pim-1, -2, and -3 kinases. Fold change was compared with 293T-pCDNA transfected controls. Luciferase activity was normalized to RL-TK renilla expression. Results represent the average of at least two experiments. (B) Verified expression of Pim kinases, -1, -2, and -3 in 293T transfected cells. Actin served as a loading control. (C) 293T cells were transfected with pcTax along with individual Pim kinases. 48-h post-transfection, gene expression was performed for Tax expression. Experiments were performed at least twice, and GAPDH served as a control. (D) 293T cells were transfected with HTLV-1 LTR luciferase construct along with pcTax and/or Pim-1 shRNA A and B. Fold change was compared with 293T-pCDNA transfected controls. Luciferase activity was normalized to RL-TK renilla expression. Results represent the average of at least two results. Western blots confirming Tax over-expression are provided. (E) 293T cells were transfected for 48 h with Tax. The pan-Pim kinase inhibitor, AZD1208 (10 μM), was added during the last 30 h. Actin served as a loading control.