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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 1. Characterization of an ER-localized redox-sensitive GFP (eroGFP).

Figure 1

(A) Fluorescence excitation spectra of recombinant fully reduced (red triangles) and oxidized (blue circles) eroGFP. Emission was measured at 520 nm. (B) Excitation spectra of recombinant eroGFP through titration of reduced to oxidized lipoic acid (ratios in inset—total 10 mM). (C) Fraction-reduced eroGFP—data from (B)—as a function of ratios of reduced to oxidized lipoic acid expressed as redox potential values. (D) Representative confocal image of cells expressing eroGFP and Sec61-mCherry. (E) eroGFP ratio (defined as the ratio of fluorescence from excitation at 488 nm vs. 405 nm expressed in log2 space and normalized to the untreated case—UT) in cells treated with the indicated concentration of DTT or H2O2. Data represent means +/− SD from three independent experiments. (F) Northern blot for HAC1 mRNA in cells expressing vector or eroGFP, treated with or without tunicamycin, Tm, (1µg/ml) for 1 hr. HAC1u denotes unspliced and HAC1i spliced mRNA respectively. An asterisk signifies the HAC1 5’ exon.