Figure 4. Expression and localization of native and episomally expressed Atg8 in P. berghei.

The trophozoite and schizont stages of wild type P. berghei were evaluated for expression and localization of native PbAtg8 using anti-Atg8 antibodies (A). Similarly, recombinant P. berghei parasites were assessed for localization of episomally expressed GFP-PfAtg8 (B) using anti-GFP antibodies and for colocalization of the signals (C) with anti-PfAtg8 and anti-GFP antibodies as described in Materials and Methods section. The images show signal for native PbAtg8 (A; Atg8) or episomally expressed GFP-PfAtg8 (B; GFP) or colocalization of anti-Atg8 and anti-GFP antibody signals (C; Atg8+GFP), staining of the nucleus (DAPI), the parasite and the erythrocyte boundaries (Bright field), and the merged of all three images (Merge). The punctate signal for both native and episomally expressed Atg8 throughout the parasite is similar to the localization pattern of native PfAtg8 in Figure 2. The yellow spots (Atg8+GFP) in C indicate that both the antibodies label the same structures and that anti-PfAtg8 antibodies are specific to PfAtg8. For representation purpose, the GFP signal was false-coloured to green and the Atg8 signal was false-coloured to red.