Figure S2.
Generation and characterization of leep1 knockout (ko) cells. (A) Design of the knockout construct. A blasticidin resistant cassette (BSR) was inserted to replace part of the open reading frame of leep1. Arrowheads mark the sites where genomic DNA was digested. (B) Targeted clones were confirmed by PCR and Southern blotting. (C) WT and leep1− cells were plated clonally with bacteria (Klebsiella aerogenes) on standard medium agar for 5 d. Scale bar = 5 mm. (D) WT and leep1− cells were plated as a monolayer on non-nutrient agar to induce development. Typical fields of view were photographed at the indicated time points. Scale bar = 2 mm. (E) Accumulation of TRITC-labeled yeast particles by phagocytosis. Scale bar = 5 µm. (F) Quantification of yeast phagocytosis. (G) Phagocytosis of K. aerogenes or Escherichia coli measured by the decreasing turbidity after addition of WT or leep1− cells. Data were from three independent experiments and represent mean ± SD. (H) Top: Trajectories of randomly migrating cells (n = 142 for WT and 168 for leep1−). Bottom: Summary of the respective motility parameters (mean ± SD). (I) Top: Trajectories of cells migrating under agarose against a passive gradient of folate (n = 155 for WT and 129 for leep1−). Bottom: Summary of the respective chemotaxis parameters (mean ± SD). Data are from three independent experiments. FMI, forward migration index.

Generation and characterization of leep1 knockout (ko) cells. (A) Design of the knockout construct. A blasticidin resistant cassette (BSR) was inserted to replace part of the open reading frame of leep1. Arrowheads mark the sites where genomic DNA was digested. (B) Targeted clones were confirmed by PCR and Southern blotting. (C) WT and leep1− cells were plated clonally with bacteria (Klebsiella aerogenes) on standard medium agar for 5 d. Scale bar = 5 mm. (D) WT and leep1− cells were plated as a monolayer on non-nutrient agar to induce development. Typical fields of view were photographed at the indicated time points. Scale bar = 2 mm. (E) Accumulation of TRITC-labeled yeast particles by phagocytosis. Scale bar = 5 µm. (F) Quantification of yeast phagocytosis. (G) Phagocytosis of K. aerogenes or Escherichia coli measured by the decreasing turbidity after addition of WT or leep1− cells. Data were from three independent experiments and represent mean ± SD. (H) Top: Trajectories of randomly migrating cells (n = 142 for WT and 168 for leep1−). Bottom: Summary of the respective motility parameters (mean ± SD). (I) Top: Trajectories of cells migrating under agarose against a passive gradient of folate (n = 155 for WT and 129 for leep1−). Bottom: Summary of the respective chemotaxis parameters (mean ± SD). Data are from three independent experiments. FMI, forward migration index.

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