Characterization of a nuclear transport factor 2-like domain-containing protein in Plasmodium berghei

Figures

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  Fig. 1. Effect of PBANKA_1019700 deletion on the asexual and sexual development of Plasmodium berghei (strain ANKA). Female C57BL/6 (B6) mice were infected with 1 × 104 erythrocytes parasitized with Plasmodium berghei ANKA with PBANKA_1019700 deletion (Δ1019700). As a control, P. berghei ANKA with p230 deletion was inoculated intravenously into mice. A Time course of parasitaemia. Results are expressed as mean ± standard deviation (SD) from six mice. Experiments using six mice were performed in triplicate. B Survival rate. Data are the mean ± SD from six mice and are representative of three independent experiments. C Cell cycle during the asexual phase. Results are mean ± SD from three independent experiments. D Bioluminescence images of luciferase activity in the organs of infected mice. For bioluminescence analysis, erythrocytes parasitized with Pb ANKA were transferred to RPMI1640 medium supplemented with 25% fetal bovine serum, 0.05 mg/mL penicillin, and 0.05 mg/mL streptomycin. The parasitized erythrocytes were incubated for 18 h under 90% N2, 5% CO2, and 5% O2. Mature schizonts and gametocytes were harvested using Nycodenz density gradient centrifugation. B6 mice were injected with…

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  Fig. 2. Effect of PBANKA_1019700 deletion on the sexual development of P. berghei ANKA. Erythrocytes parasitized with Plasmodium were incubated for 22 h, 28 h, and 45 h. The percentages of male and female gametocytes were calculated as follows: ([number of male or female gametocytes] ÷ [total number of schizonts plus male and female gametocytes]) × 100. A Percentages of mature female gametocytes. B Percentages of mature male gametocytes. Results are expressed as the mean ± SD from three independent experiments. *Significant difference compared to controls

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  Fig. 3. Live-cell fluorescence imaging of 1019700::mCherry-expressing parasites. Female B6 mice were infected with 1 × 104 erythrocytes parasitized with transgenic Plasmodium expressing mCherry fused to PBANKA_1019700 (red). The cells’ nuclei were stained with Hoechst 33342 (blue). At least 50 parasitized erythrocytes were analysed, and the same fluorescence pattern was observed in all parasitized erythrocytes. Representative data are shown. Scale bar = 10 μm

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  Fig. 4. Effect of PBANKA_0519900 deletion on the asexual and sexual development of P. berghei ANKA. Female B6 mice were infected with 1 × 104 erythrocytes parasitized with P. berghei ANKA with PBANKA_0519900 deletion (Δ0519900). As a control, P. berghei ANKA with p230 deletion was inoculated intravenously into mice. *Significant difference compared to controls. A Time course of parasitaemia. Results are expressed as mean ± SD from four mice. Experiments using four mice were performed in triplicate. B Survival rate. Data are the mean ± SD from four mice and are representative of three independent experiments. C, D Percentages of mature female (C) and male (D) gametocytes. Erythrocytes parasitized with Plasmodium were incubated for 28 h. The percentages of male and female gametocytes were calculated as follows: ([number of male or female gametocytes] ÷ [total number of schizonts plus male and female gametocytes]) × 100. Results are expressed as the mean ± SD of three independent experiments

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  Fig. 5. Cellular localization of PBANKA_0519900 and PBANKA_1359300 in P. berghei ANKA. Female B6 mice were infected with 1 × 104 erythrocytes parasitized with transgenic Plasmodium expressing 0519900::GFP and 1019700::mCherry, or transgenic Plasmodium expressing 1359300::GFP and 1019700::mCherry. The cells’ nuclei were stained with Hoechst 33342 (blue). At least 50 parasitized erythrocytes were analysed, and the same fluorescence pattern was observed in trophozoites. A Transgenic parasites expressing 0519900::GFP (0519900, green) and 1019700::mCherry (1019700, red) during the trophozoite stage are shown. B Transgenic parasites expressing 1019700::mCherry (1019700, red) and 1359300::GFP (1359300, green) during the trophozoite stage are shown. Representative data are shown. Scale bar = 10 μm