Fig. 1. GFP-Yck2p induction results in accumulation of intracellular punctate fluorescence before localization to the plasma membrane. (A) Induction of GFP-Yck2p was monitored by immunoblot analysis (Panek et al., 1997) using an affinity-purified antiserum against GFP. Detection in extracts from glucose-grown cells is shown in the left panels, for pJB1 (pGal:GFP:YCK2; lane 1) in wild-type strain LRB939, for GFP-Yck2p expressed from a chromosomal copy under the control of the YCK2 promoter in strain LRB913 (lane 2), and from a high copy (2µ) plasmid (pL2.35; lane 3) in LRB939. For the right panels, cells were grown at 30°C in synthetic medium containing 2% raffinose (derepressing conditions) and galactose was added to 2% at the 0 minute time point. Extracts were prepared from culture aliquots taken at the time points indicated. Antiserum against phosphoglycerate kinase (PGK) was used to reprobe the blots for loading control. (B) DIC and fluorescence images of living cells treated as in B were captured at the indicated time points. Bar, 2 µm.

Return to article