Fig. 6

Expression pattern of Glγ-giardin in G. lamblia during encystation. a Intracellular level of Glγ-giardin during G. lamblia encystation was performed by western blot analysis. Giardia extracts were prepared at various time-points in trophozoites (0) and encysting cells (6, 12, 24 and 48 h post-induction to encystation). Immunoreactive bands were observed at 35 or 21 kDa, expected sizes of Glγ-giardin or GlCWP1, respectively. After deprobing antibodies with stripping buffer, membrane was incubated with polyclonal rat antibodies specific to PDI1 of G. lamblia (1:10,000) as loading control. An immunoreactive band was detected at 26 kDa. Relative expression of Glγ-giardin during encystation is presented as a bar graph. b Localization of Glγ-giardin in HA-tagged Glγ-giardin expressing G. lamblia during encystation. Giardia lamblia attached to glass slides were reacted overnight with mouse anti-HA (1:100) and rat anti-GlCWP1 polyclonal antibodies (1:400) then incubated with Alexafluor 488-conjugated anti-mouse IgG (1:100) and Alexafluor 555-conjugated anti-rat IgG (1:200). Cells were mounted with DAPI containing anti-fade mounting medium and observed. DIC image was acquired to show cell morphology. Scale-bars: 2 μm