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. Author manuscript; available in PMC: 2014 Oct 1.
Published in final edited form as: Nat Methods. 2014 Mar 9;11(4):443–448. doi: 10.1038/nmeth.2879

Figure 2. The automated FACS technology allows for facile and reproducible growth rate-corrected determination of reporter protein synthesis rate, and degradation fluxes as a function of time.

Figure 2

(a) Instantaneous cell division rate of cells stressed with tunicamycin (Tm). Cell division rates decrease in a dose-dependent way but (b) the average volume increases. (c) Average fluorescence measurement of the p4XUPRE-GFP transcriptional reporter, (d) Growth corrected expression rate of p4XUPRE-GFP. (e) Average fluorescence of pTDH3-mKate2. TDH3 is a housekeeping gene (f) Growth corrected expression rate of pTDH3. (g) Distributions of raw fluorescence across time for a mixture of two strains in a competition experiment where they are distinguished by the presence or absence of a pTDH3-mKate2 fluorescent tag (left). Distributions of green fluorescence in the two strains, containing GFP (red population) or a short-lived allele Ub-Tyr-GFP (blue population), both driven by the UPR synthetic promoter 4XUPRE (right). (h) Raw average fluorescence for the unstable GFP allele for different doses of ER stress induced by addition to tunicamycin. (i) Differential degradation flux of the unstable allele Ub-Tyr-GFP as a function of time. Experiment was replicated twice in the laboratory.