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Figure 2


Fig. 2. Subcellular localizations of TLK1 and TLK2 in T. brucei. (A) Cells expressing HA-tagged wild-type and kinase-dead TLK1 and TLK2 were boiled in SDS sample solution and immunoblotted with mAb against (`{alpha}') HA and mAb against {alpha}-tubulin. (B) Cell lysates were dialyzed and incubated with (+ {lambda}PPase) or without (– {lambda}PPase) lambda protein phosphatase, and blotted with mAb against HA and mAb against {alpha}-tubulin. (C) HA-tagged TLK1 and TLK2 were expressed in 29-13 (WT) cells or in cells harboring the AUK1 RNAi construct (AUK1 RNAi). Lysates from un-induced (– Tet) and tetracycline-induced (+ Tet) cells were immunoblotted with mAb against HA and mAb against {alpha}-tubulin. (D) Subcellular localizations of wild-type and kinase-dead TLK1 and TLK2. Cells were co-stained with KMX-1 antibody for the spindle (red) and FITC-conjugated antibody against HA for HA-tagged proteins (green) and DAPI for DNA (blue). Arrows point to the brightly stained TLK1 spots, which correspond to the positions of spindle poles. (E) Subcellular localization of TLK1-HA in the AUK1 RNAi cell. The cell was co-stained with KMX-1 and antibodies against HA and counterstained with DAPI. The arrow points to a focal point of TLK1 inside the nucleus. Bars, 2 µm.