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. 2017 Mar 1;8(15):24785–24796. doi: 10.18632/oncotarget.15806

Figure 2. Expression of ectopic GPC3 by stably transfected P. yoelii 17XNL.

Figure 2

(A) Schematic representation of the pL0017-gpc3 vector integrated into the c-rrna unit. (B) Gel electrophoresis and DNA analysis of the gpc3 cloned from cDNAs of Hepa1-6 cells (left) by PCR, and identification of the pL0017-gpc3 vector (right) by restriction enzyme analysis (Bam HI/Xba I). (C) Correct integration of the vector in P.y-GPC3. PCR was performed with genomic DNA from P.y-GPC3 and P.y-WT (negative control). The 5′ integration site (5′ int, primers PyL739/L635) and 3′ integration site (3′ int, primers L665/PyL740) were verified, as well as the gpc3 gene. (D) Detection of GPC3 expression in P.y-GPC3 by western blot using anti-GPC3 (rabbit) or anti-Flag tag antibodies. (E) GPC3 expression by the four different stages of the P.y-GPC3. Parasitized erythrocytes were analyzed by immunofluorescence and P.y-WT infected erythrocytes were used for the negative control. Parasites stained with anti-GPC3 antibody (green), or DAPI for DNA, were visualized by confocal microscopy. Bars: 5 μm.