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. Author manuscript; available in PMC: 2015 Sep 29.
Published in final edited form as: Dev Cell. 2014 Sep 11;30(6):759–767. doi: 10.1016/j.devcel.2014.07.016

Figure 2. Overexpression of Ypt1 and Ypt31 rescues Sec7 allele-specific phenotypes.

Figure 2

(A) Localization of GFP-tagged Sec7, sec7-1, and sec7-4 after 20 min incubation at restrictive temperature (37°C).

(B) Localization of GFP-sec7-1 in cells overexpressing Ypt1 at both permissive (30°C) and restrictive (35°C) temperatures.

(C) Activation of Arf1 (measured via tryptophan fluorescence) by 100 nM WT Sec7f, sec7-4f, or sec7-4f in the presence of 500 nM activated Ypt31. Dark lines represent the average of 3 normalized reactions; lighter shaded areas represent the corresponding 95% confidence intervals (CI); dashed lines represent data not captured but inferred from curve-fitting.

(D) Quantification of Arf1 activation rates from curves in (C). Data for WT Sec7f + Ypt31 is included for comparison. The activation curve for this sample is not shown in (C) because it was acquired using a different concentration (30 nM) of the GEF. Error bars represent 95% CIs for n=3.