Flow cytometry analysis was performed using Gal1-GFP on HTZ1/htz1Δ (JRY9007), htz1-K3,8,10,14R/htz1Δ (JRY9008), htz1-K3R/htz1Δ (JRY9009), htz1-K8R/htz1Δ (JRY2010), htz1-K10R/htz1Δ (JRY2011), and htz1-K14R/htz1Δ (JRY2012) cells grown long-term in YP-glucose (2%) prior to being transferred into YP-galactose (2%). A threshold level of GFP-intensity was set so that between 1% and 2% of glucose-grown HTZ1 cultures were classified as GFP-positive cells. (A) The frequency of GFP-positive cells within HTZ1, htz1-K3,8,10,14R, htz1-K3R, htz1-K8R, htz1-K10R, and htz1-K14R cultures. (B) The average GFP intensity of the GFP-positive populations of HTZ1, htz1-K3,8,10,14R, htz1-K3R, htz1-K8R, htz1-K10R, and htz1-K14R cultures. Bars in all panels represent the standard deviations of values from three biological replicates.