Fig 4.

ROB1 in the SC5314 background is heterozygous and the two alleles have different filamentation phenotypes. (A) Six independent isolates of heterozygous ROB1 strains derived in the SN background were plated on solid Spider medium and incubated for 4 days at 37°C. (B) Diagrams of the Rob1 ORF indicating the position of the SNP and the Sanger sequences of the six heterozygous strains shown in (A). (C) The distribution of cell morphologies after 4 hours incubation in RPMI + 1% BCS. The asterisk indicates that the indicated strain differs from SN250 in a statistically significant manner by Student’s t test (P <0.05). (D) Micrographs of mouse ears inoculated with a 1:1 mixture of SN250 (GFP labeled) and the indicated rob1 mutant (RFP) and imaged by confocal microscopy 24 hours post infection. Quantitation of the percentage of filamentous cells for the two strains (E and F). For details of imaging and quantitation see Materials and Methods. NS indicates the differences were not significant by Student’s t test (P > 0.05).