LmjF.21.0725 is essential for <i>L</i>. <i>major</i> promastigotes.

(A) FACS analysis was used to quantify GFP levels and estimate dynamics of passive loss of pXNeoGFP-LmjF.21.0725 in four independent transfectants in the absence of G418(depicted as numbers 1 to 4, top of the panel). (B) With same culture conditions, western blot analysis indicated differences in LmjF.21.0725 expression levels in transfectant lines, using anti-LmjF.21.0725 antibody. (C) Fluorescent microscopy images showing GFP expression in transfectants (10th passage in absence of G418). Lines: +/PAC (+) = heterozygous cell line with pXNeoGFP-LmjF21.0725; HYG/PAC(+) = double replacement transfectant bearing both selectable markers and plasmid expressing LmjF.21.0725 (pxNeoGFP-LmjF21.0725, addback transfectant).