Additional file 2: of Using in vivo oxidation status of one- and two-component redox relays to determine H2O2 levels linked to signaling and toxicity

Figure S2. Kinetics of OxyR and Tpx1 oxidation. a In cells deficient in Tpx1 or Trx1, HA-OxyR oxidizes at low concentrations of peroxides. Aerobic or anaerobic cultures of strains AD29 (WT), AD61 (Δtrx1), and AD36 (Δtpx1) carrying an integrative sty1 promoter-driven HA-oxyR gene were treated or not with the indicated concentrations of H2O2 for the times indicated. TCA extracts were analyzed as in Fig. 2a. b Cultures of strain AD29 (WT) were treated or not with 20, 50, or 100 μM H2O2 for the times indicated. TCA extracts were analyzed as in Fig. 2a, using antibodies against Tpx1 (ox. Tpx1 dimer is the upper band in the panels; red. Tpx1 monomer is the lower band in the panels). (PDF 476 kb)