Fig. 2

Construction of transgenic Giardia trophozoites expressing HA-tagged Glγ-giardin and localization of HA-tagged Glγ-giardin. a Expression of HA-tagged Glγ-giardin was confirmed by western blot analysis. Extracts were prepared from G. lamblia containing pΔ.pac (Lane 1), or pGlγ-giardinHAX3.pac (Lane 2). Membrane was reacted with monoclonal mouse anti-HA (1:1000). 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. b Localization of Glγ-giardin in G. lamblia expressing HA-tagged Glγ-giardin. Giardia lamblia expressing HA-tagged Glγ-giardin attached to glass slides were reacted overnight with mouse anti-HA (1:100) and anti-Glγ-giardin (1:100) then incubated with Alexafluor 488-conjugated anti-mouse IgG (1:100) and Alexafluor 555-conjugated anti-rat IgG (1:100). DIC image was acquired to show cell morphology. Scale-bars: 2 μm. c Percentages of Giardia cells with stained MB at G1/S- and G2-phases