Fig. 5.
PRD-1 is nucleus-localized in glucose sufficient cultures but is lost upon glucose starvation. (A) Mycelia expressing PRD-1GFP were grown in 2% (wt/vol) glucose liquid medium overnight and transferred to liquid medium containing either 2% (wt/vol) or 0% glucose. Mycelia were fixed and stained with AlexaFluor 488-conjugated GFP antibodies at the given time points. PRD-1 is shown in yellow, and nuclei are shown by Hoechst in cyan. All images were collected with a 63× objective. (Scale bars: 10 µm.) Levels of glucose in the medium were confirmed using the Sigma Glucose (GO) Assay; after transfer to 0% glucose medium, average soluble glucose at the five times noted was 0.0128 ± 0.006 mg/mL SEM (n = 5) and was 0.00 at 10 h whereas glucose remained above 16 mg/mL in the glucose replete cultures. (B) Mycelia were grown in 0.1% glucose liquid medium for 16 h overnight; glucose was used so that measured available glucose was 0.000 mg/mL Mycelia were then transferred to flasks containing 2% (wt/vol) glucose and sampled at the times noted.