Fig. 1: Overview of putative Leishmania bromodomain factors.

a Schematic showing protein domain architecture of Leishmania BDFs. b Overview of Cas9 gene deletion attempts of BDF1–7 in L. mexicana T7/Cas9 promastigotes. Two independent transfections were carried out, one using only BSD, another using both BSD or NEO as a drug selectable markers. For the BSD plus NEO experiment clones were selected using blasticidin and G418 in combination, or individually. No clones were recovered from the dual selection, but null mutants of BDF6 and BDF7 were recovered on all 3 occasions when a single drug selection was applied. No null mutants of BDF1–5 were ever recovered. c Live-cell fluorescent microscopy of L. mexicana promastigote expressing mNG::BDF5. Nucleus is denoted by arrowhead labelled n, the kinetoplastid DNA is indicated by arrowhead labelled k. d Channel separated Z-slices of the nucleus from the cell in (c). e Live-cell fluorescent microscopy of intramacrophage L. mexicana amastigotes expressing mNG::BDF5 endogenously tagged protein. f Expression levels of mNG::BDF5 during promastigote growth, determined by mNG signal in individual cells by flow cytometry. Dashed points denote mean cell density, error bars ± standard deviation, solid points denote median fluorescence intensity, N = 3,20,000 events per sample.