Figure 6
Transient expression of Ty-tagged fluorescent eGFP protein. (A) The plasmid used for ectopic, transient expression of eGFP in Blastocystis was driven by the legumain promoter and 3′ UTR. (B) A total of 108 cells were transfected with 100 μg of plasmid DNA, using “Time Constant” protocol (1 pulse of 370 V, 20 ms). The transfectants were washed with PBS and fixed with 4% PFA. After permeabilization with 1% NP-40 in PBS, the cells were blocked with 3% BSA, and stained with mouse-anti-Ty monoclonal primary antibody followed by goat-anti-mouse IgG (Alexa Fluo 594) secondary antibody. The eGFP and Ty signals were monitored using fluorescent microscopy. Scale bar, 10 μm.
