FIG 5.

Pim kinases bind Tax. (A) 293T cells were co-transfected with pcTax and PMH-Pim kinases for 48 h. Lysates were subject to co-immunoprecipitation with HA antibody and probed with anti-Tax and anti-HA antibodies. Whole cell lysates were probed for Tax and HA-Pim expression to verify expression. Actin demonstrates the amount of protein in each lysate. (B) 293T cells were transfected with pcTax and HA-tagged Pim-1 expression plasmids for 48 h. Cells were treated with or without proteosome inhibitor, MG132 (10 μM) for the last 8 h. (C) 293T cells were transfected with pcTax and HA-tagged Pim-1 expression plasmids for 48 h. Cells were treated with or without proteosome inhibitors, MG132 (10 μM) and lactacystin (10 μM) for the last 8 h. (D–F) 293T cells were transfected with pcTax and HA-tagged Pim-1, -2, or-3 expression plasmids for 48 h. Cells were treated with or without proteosome inhibitor, MG132 (10 μM), or lysosomal inhibitors, chloroquine (100 μM) or leupeptin (100 μM) for the last 8 h. For (B–F), whole cell lysates were probed with Tax and anti-HA (Pim) antibodies. Actin served as a loading control.