Figure 3

High-throughput stable transfection. Three plasmids, pfGNr (green), pHHT-TK-Δrh3 (black), pCC1-Δrh3 (blue) were transfected using the optimized protocol (see Methods). Dots on the growth line represent days at which wells were screened for parasites either through fluorescence microscopy (pfGNr) or Giemsa-stained smears (pfGNr, pCC1-Δrh3 and pHHT-TK-Δrh3). Schedule of outgrowth conditions consists of weekly blood additions (0.5% HC, light red drop) and 2:3 culture splits (red drop). Note a large split of the culture at two weeks post-transfection. Data are shown as percentage positive wells where the detection of a ring in a culture well equals a positive well and 24 and 16 are the total number of wells for pfGNr and for pCC1 and pHHT-TK, respectively. Day 0 = transfection day.