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. 2016 Aug 2;19(2):e12646. doi: 10.1111/cmi.12646

Figure 4.

Figure 4

Chemical inhibition of GLUT1‐mediated glucose transport by WZB117 affects P. berghei hepatic infection in vitro (A‐C) and in vivo (D‐G). A. Huh7 cells were infected with luciferase‐expressing P. berghei sporozoites and 2 h later the culture medium was replaced by medium with increasing concentrations of WZB117. Parasite load (luminescence) and cell viability were assessed at 48 hpi. Representative experiment out of two independent experiments. Error bars represent standard deviation (SD). One‐way analysis of variance (ANOVA) with post‐test Dunnett. B. Huh7 cells were infected with red fluorescent protein‐expressing P. berghei sporozoites and 2 h later the culture medium was replaced by medium with increasing concentrations of WZB117. Parasite development was assessed by flow cytometry at 48 hpi. Pool of two independent experiments. Error bars represent SD. One‐way ANOVA with post‐test Dunnett. C. 2‐NBDG uptake by red fluorescent protein‐expressing P. berghei‐infected cells at 48 hpi after treatment with increasing concentrations of WZB117, assessed by flow cytometry. Pool of two independent experiments. One‐way ANOVA with post‐test Dunnett. D. C57BL/6 mice received three doses of 10 mg/kg of WZB117 i.p., the first immediately before infection with P. berghei sporozoites and the other two at 15 and 30 hpi. Parasite load was assessed at 44 hpi by qPCR. Pool of three independent experiments. Vehicle: n = 15 mice; WZB117‐treated: n = 12 mice. Two‐tailed Mann–Whitney test. E. Fifty µm sections of one liver lobe from one mouse of each experimental group were stained with anti‐UIS4 (white), anti‐GFP (green), phalloidin (red) and Hoechst (blue). Representative confocal images of P. berghei parasites in both experimental groups of mice. Scale bar, 10 µm. F. Area of the parasites in the control (vehicle) or the WZB117‐treated mouse, determined by immunofluorescence microscopy. Representative experiment out of two independent experiments. Two‐tailed Mann–Whitney test. G. Number of parasites per area of liver in control and WZB117‐treated mice. Representative experiment out of two independent experiments. Error bars represent SD. Two‐tailed Mann–Whitney test. ns, not significant; * P < 0.05, ** P < 0.01 and *** P < 0.001.