FIG 8.

Inhibition of Pim kinases leads to elevation of Tax expression in HTLV-1 lines. (A–C) Treatment of MT1, MT2, ATL25 (A), and ATLT (B–C) with 5, 10, or 20 μM AZD1208, or 10 μM M-100 or PimIV inhibitor. Treatments were performed for 48 h or 72 h. Representation of at least two independent experiments is shown. Gene expression was normalized to GAPDH expression and performed at least twice. (D–E) Jurkat TET-Tax cells were induced with doxycycline for 72 h. 10 μM AZD1208 was added for the final 18 h for gene expression (D) or 72 h for protein expression (E). Actin served as a loading control. (F) ATLT and MT4 cells were treated with 10 μM AZD1208 and/or 10 μM MG132. Total cell lysate was probed for Tax and actin served as loading control. (G) ATLT and MT4 were treated with 10 μM AZD1208. Cytoplasmic, soluble nuclear, and insoluble nuclear fractions were probed with Tax antibody. Purity of fractions was confirmed with Cyclin B1 and α-Tubulin antibodies.