Fig. 3

Verification of the expression of the GP40 and GP15ΔGPI of Cryptosporidium parvum in Toxoplasma gondii by immunofluorescence assay. a CpGP40 and CpGP15ΔGPI expressed in T. gondii. T. gondii RHΔku80 tachyzoites were transfected with pGRA1-CpGP40 or pGRA1-CpGP15Δgpi and allowed to infect HFF monolayers. IFA was performed 24 h after infection with anti-GP60 and -HA followed by Alexa Fluor-488-conjugated and 594-conjugated secondary antibodies. The far left panel shows the DIC field, and the far right panel shows a merged fluorescence image of the same field. DAPI-stained nuclei are in blue, Alexa Fluor-488 is in green, and Alexa Fluor-594 is in red. b CpGP40 expressed in T. gondii. IFA was performed 24 h after infection with anti-GP60 and -Strep followed by Alexa Fluor-488-conjugated and 594-conjugated secondary antibodies. c High-power images of the IFA to show the fine localization of the GP40 expression. CpGP40 proteins expressed in T. gondii are localized to the parasitophorous vacuole. Scale bars = 5 μm