Figure 7. Efr3 is a component of PIK... | Rockefeller University Press

Figure 7.

$Figure 7. Efr3 is a component of PIK patches at the PM. (A) Tetrad dissection of diploid yeast with a deleted chromosomal copy of efr3. (B) Full-length and truncation mutants of GST-tagged Efr3 were purified from E. coli lysates, immobilized on glutathione beads, and normalized to equal amounts. Beads containing the GST-Efr3 constructs were then incubated for 1 h with yeast lysate expressing Ypp1-GFP. Protein that bound the GST-Efr3 fragments was probed using an anti-GFP antibody. (C) Subcellular fractionation of Efr3-GFP from wild-type cell extracts. (D) Cells coexpressing Efr3-GFP and Ypp1-mCherry were grown at 26°C and examined by fluorescence microscopy. Efr3-GFP localizes to cortical patches on the PM (left) and with a similar distribution as Ypp1-mCherry (right). Arrows on the Efr3-GFP image indicate regions of colocalization. Bar, 4 μm.$

Efr3 is a component of PIK patches at the PM. (A) Tetrad dissection of diploid yeast with a deleted chromosomal copy of efr3. (B) Full-length and truncation mutants of GST-tagged Efr3 were purified from E. coli lysates, immobilized on glutathione beads, and normalized to equal amounts. Beads containing the GST-Efr3 constructs were then incubated for 1 h with yeast lysate expressing Ypp1-GFP. Protein that bound the GST-Efr3 fragments was probed using an anti-GFP antibody. (C) Subcellular fractionation of Efr3-GFP from wild-type cell extracts. (D) Cells coexpressing Efr3-GFP and Ypp1-mCherry were grown at 26°C and examined by fluorescence microscopy. Efr3-GFP localizes to cortical patches on the PM (left) and with a similar distribution as Ypp1-mCherry (right). Arrows on the Efr3-GFP image indicate regions of colocalization. Bar, 4 μm.