Cell - 8 September 2016

Figure S3. Construction and Validation of the Cas9 Strain Expressing H2B-YFP, Related toFigure 5
(A) Diagram of the constructs used to generate theT. gondiistrain expressing H2B-YFP and Cas9. The sequence of the decoy sgRNA is highlighted in the top
construct, followed by the Cas9-binding sequence (orange).
(B) Expression of H2B-YFP in the nucleus of the clonal parasites detected by live fluorescence video microscopy.
(C) Representative micrographs showing intracellular parasites three days post transfection. Parasites were stained for SAG1 (red), and TgACT1 (ACT; green).
Host-cell and parasite nuclei were stained with DAPI (blue). Scale bar, 10mm. SAG1 retention in wild-type (wt) and Cas9-expressing parasites was measured
following the different treatments.