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. Author manuscript; available in PMC: 2009 Nov 28.
Published in final edited form as: Cell. 2008 Nov 20;135(5):933–947. doi: 10.1016/j.cell.2008.10.011

Figure 2. Dynamic monitoring of ER redox status and UPR activity during pharmacologically-induced ER stress.

Figure 2

(A) Schematic of composite reporter gene. (B) Schematic showing configuration of flow cytometer laser lines and filters. A syringe-pump periodically (~10 minutes) injected 25µl of sample from cultures growing in bioreactors into a flow cytometer. (C–F) Time courses of eroGFP ratio or UPR-RFP metric histograms in wild-type cells during treatment with DTT (2mM) or Tm (1µg/ml). The eroGFP ratio and UPR-RFP metrics are normalized to wild-type unstressed cells. Color represents percentage of cells at a given metric value and time point. Dashed gray line signifies time of DTT or Tm addition. (G) Non-reducing SDS-PAGE and immunoblot (anti-GFP) of AMS-treated protein extracts from untreated cells (lane 1), cells treated with 10mM DTT for 30 min (lane 2), or 2µg/ml Tm for 3 hrs (lane 3).