Fig. 1.

Transfection of pleomorphic transfections from cells harvested from blood or from culture. (A) Diagram outlining the method for the harvest and transfection of pleomorphic slender forms (details described in main text). (B) Pleomorphic bloodstream form AnTat1.1 90:13 cells were harvested from mouse blood 3 days post infection or after 7 days in vitro. Between 2.8 and 4.06 × 10⁷ cells were transfected with 10 μg of pALC14 vector in the “Amaxa basic parasite nucleofector solution 2” transfection buffer and selected in 24 well plates at 1:2, 1:5, 1:25 and 1:125 dilutions with 0.5 μg/ml puromycin. Transfection efficiencies were calculated from the number of positive wells per dilution (excluding any dilution where >50% of wells were positive) and extrapolating for the total number of cells transfected. Replicates were carried out from three independent mouse infections and three independent in vitro cultures.