Figure 2.

H₂O₂-dependent oxidation of roGFP2-Tpx1.C169S expressed in S. pombe, Jurkat T cells, and S. cerevisiae. Wild-type of S. pombe (HM123), S. cerevisiae (BY4741), or Jurkat T cells, transformed with p407.C169S, p791, or p797 to express roGFP2-Tpx1.C169S, were grown in their own media (MM-, SD-, or RPMI 1640-supplemented), centrifuged and resuspended in MM (both yeasts) or ISO (Jurkat) for a final OD₆₀₀ of 1 (both yeasts) or 0.5 (Jurkat cells). Then, 190 µL of these cell suspensions were plated in 96-well imaging plates, and incubation and fluorescence recording were initiated at 37 °C with 100 rpm shaking. After 4 cycles, the indicated treatments of DTT or H₂O₂ were applied (indicated with arrows). The degree of probe oxidation, or OxD (amount of probe oxidation per 1), is indicated over time. (b) Bar graphs represent the average of 3 OxD values from the 12 to 17 min time points for each treatment and cell type represented in (a). Statistical significance was calculated between the indicated samples with a ratio paired Student’s t-test with p-values of 0.05 (*) and 0.001 (***); n.s., non-significant. For each strain, average data from four biological replicates are shown, with error bars (S.D.) of (a) displayed in Figure S1.