Figure S4.
Characterization of ras knockout cells. (A) Localization of GFP-tagged RasB, RasD, and RasG in WT cells. Scale bar, 5 μm. (B) Time-lapse imaging of Byr2RBD-GFP in WT cells during macropinocytosis. Scale bar, 5 μm. (C) Top: Design of ras knockout construct. A blasticidin-resistant cassette (BSR) or a Hygromycin-resistant cassette (HygR) was inserted to replace part of the open reading frame of rasB, rasD, or rasG via homologous recombination. Bottom: Knockout clones were confirmed by PCR using the indicated primers. (D) Growth of WT and ras knockout cells on bacterial lawns. Cells were plated clonally with bacteria (Klebsiella aerogenes) on standard medium agar for 5 days. Scale bar, 2 mm. (E) Generation time of WT and ras knockout cells. The plot shows means and SEM; data were from three independent experiments. (F) Localization of GFP-Leep2A and RFP-Leep2B in WT and ras knockout cells. Scale bar, 5 μm.

Characterization of ras knockout cells. (A) Localization of GFP-tagged RasB, RasD, and RasG in WT cells. Scale bar, 5 μm. (B) Time-lapse imaging of Byr2RBD-GFP in WT cells during macropinocytosis. Scale bar, 5 μm. (C) Top: Design of ras knockout construct. A blasticidin-resistant cassette (BSR) or a Hygromycin-resistant cassette (HygR) was inserted to replace part of the open reading frame of rasB, rasD, or rasG via homologous recombination. Bottom: Knockout clones were confirmed by PCR using the indicated primers. (D) Growth of WT and ras knockout cells on bacterial lawns. Cells were plated clonally with bacteria (Klebsiella aerogenes) on standard medium agar for 5 days. Scale bar, 2 mm. (E) Generation time of WT and ras knockout cells. The plot shows means and SEM; data were from three independent experiments. (F) Localization of GFP-Leep2A and RFP-Leep2B in WT and ras knockout cells. Scale bar, 5 μm.

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