Fig. 6

Expression and localization of phosphorylated GlPLK in G. lamblia. a Giardia lamblia cells expressing HA-tagged GlPLK in the interphase (lane 1), G1/S phase (lane 2), and G2/M phase (lane 3). Western blot analysis. Extracts of these cells were probed with anti-phospho-PLK, anti-HA, anti-GlPLK or anti-GlPDI1 antibodies. Levels of phospho-GlPLK-HA, phospho-GlPLK, GlPLK-HA and GlPLK were normalized to GlPDI1, a protein loading control. Amounts of these GlPLK proteins are expressed as relative values to those observed in the interphase cells. The presented western blot is a representative of three independent experiments, and averages of three experiments are presented as a bar graph. b Localization of GlPLK and phosphorylated GlPLK in Giardia. G. lamblia expressing HA-tagged GlPLK were incubated with antibodies specific to the phosphorylated form of PLK (1:100) along with anti-HA antibodies. These cells were then reacted with anti-Alexa Fluor 568-conjugated anti-mouse IgG and Alexa Fluor 488-conjugated anti-rat IgG. Slides were mounted with ProLong™ Gold Antifade Mountant with DAPI, and then examined with a Zeiss LSM700 inverted confocal laser scanning microscope. Scale bars: 2 µm