Localization of CRK9 in procyclic cells. CRK9 fused to the 3xHA tag was inserted into the pc-BLA-PTP vector. After linearization, transfection into T. brucei, and selection using blasticidin (10 μg/ml), cells expressing CRK9-3HA at the endogenous level were obtained. Transfected and control cells were analyzed by Western blotting for the HA tag as indicated (A). The blot was also probed with anti α-tubulin to verify equal sample loading. Localization of CRK9 was determined by fixing the transfected cells and staining them with a FITC-conjugated anti-HA antibody (B). DAPI was used to visualize the nucleus and kinetoplast and merged images were created using the imageJ software.
Gourguechon and Wang BMC Cell Biology 2009 10:68 doi:10.1186/1471-2121-10-68