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. 2014 Feb 25;9(2):e90072. doi: 10.1371/journal.pone.0090072

Figure 3. The plant AtSec24A ameliorates the phenotypes displayed by the yeast sec24-11 mutant.

Figure 3

A) The wild-type (WT) cells and the sec24-11 mutant transformed with empty vector (pVV) or expressing AtSEC23/24L, AtSEC24A and AtLST1A and B were grown at 25°C to mid-exponential phase prior to be spotted on YPD medium containing MATa bar1 mutant cells to determine α-factor secretion at permissive (25°C) and various restrictive temperatures (30°C and 37°C). B) The sec24-11 mutant cells transformed with either pVV or AtSEC24A plasmids and expressing GFP-Snc1 were grown at 25°C to mid-exponential phase, then half of cell cultures was shifted at 25°C or 37°C for 2 h prior to their observation by fluorescence microscopy to detect the intracellular localization of the GFP-Snc1 SNARE. C) Total proteins were extracted from the same strains as in B) but after a 6 h shift at 37°C and a western-blot with anti-GFP antibodies was done to detect the state of phosphorylation of the GFP-Snc1 proteins.