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same color coding as for Figure 1.) and PCR analysis to select edited clones using primers from the 5' side of PF16 (AH62:AH63) and 3' side (AH64:AH65); from left to right: WT, PS and clone 1 (cl1): one tested clone showing 100% edition rate (no WT allele detected on both the 5' and 3' sides). (B) PCR analysis performed on genomic DNA extracted at different time points post-induction using PF16-specific primers: a decrease in PF16 amplification was observed after induction, followed by a complete extinction of the signal at day 8-10 post-induction. (C) Immunofluorescence of a PF16-HA clone (100% edited) using an anti-HA antibody before (Non-induced) and after (Induced) rapamycin induction. (D) PF16 signal quantification based on immunofluorescence performed at different time points post-induction; >100 parasites were counted for each time point and classified as positive or negative labeling, Leishmania Supplementary Figure 3: PF16 mutants as another proof of concept for the Floxed gene strategy (A) Scheme of the edited locus of interest an HA tag was positioned on the 3' side (top ,