Ms excel 2013

For the analysis of macrogamete and zygote formation, mature gametocytes of WT NF54, the 7-Helix-1-KO lines 2E6 and 1D12 and the complementation line 7-Helix-1-KO(+) were enriched via Percoll purification and set to a gametocytemia of 2–4%. For inhibitor experiments, the parasites were incubated with emetine or cycloheximide at IC₅₀ concentration (as determined by Malstat assay) for 30 min at 37°C. Incubation with the solvents alone was used as a negative control. The gametocytes were activated and at 30 min p.a. (macrogametes) or 4 h p.a. (zygotes), the samples were subjected to IFA. Immunolabeling was performed as described above and macrogametes and zygotes were labelled with rabbit anti-Pfs25 antisera. Parasites were counted microscopically using a Leica DM 5500B fluorescence microscope with 600-fold magnification. For comparative macrogamete and zygote formation assays, 30 optical fields were counted for three to four times. The relative numbers of macrogametes and zygotes were calculated (WT set to 100%). Three to eight independent experiments were conducted; data analysis was performed using MS Excel 2013 and GraphPad Prism 5. For inhibitor experiments, macrogametes and zygotes per 1,000 RBC were counted for five times. Two independent experiments were performed; data analysis was performed using MS Excel 2013 and GraphPad Prism 5.