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. Author manuscript; available in PMC: 2019 Jan 10.
Published in final edited form as: Sci Signal. 2018 Jul 10;11(538):eaaq0702. doi: 10.1126/scisignal.aaq0702

Figure 4. Thioredoxin 1 negatively regulates ATM redox dimerization.

Figure 4.

A, Western blot analysis of HeLa cells expressing shRNA targeting EGFP (shEGFP) as a negative control or thioredoxin 1 (shTRX1) treated with vehicle or the indicated concentration of MD for 30 minutes. Blot was probed for ATM monomer (ATM-M), ATM dimer (ATM-D), and TRX1, with calnexin as a loading control. Blots are representative of two experiments. B, DCFDA staining by flow cytometry, in cells treated with vehicle, 5 μM or 20 μM MD as described in A. Plot is representative of three experiments utilizing biological duplicate samples in each. C, Western blot of HeLa cell treated with 20 μM MD for 30 minutes, after which the media was changed and cells were allowed to recover for 0, 10, 20, 30, 40 minutes as indicated. Blot was probed for ATM monomer (ATM-M), ATM dimer (ATM-D) and TRX1, with HSP60 and GAPDH as loading controls. Blots are representative of three experiments. D, Western blot of HeLa cells transfected with control or siRNA targeting PRDX1 and PRDX2 for 72 hours and treated with vehicle (−) or 20 μM MD (+) for 30 minutes. Blot was probed for ATM monomer (ATM-M), ATM dimer (ATM-D), PRDX1 and PRDX2, with calnexin as a loading control. Blots are representative of three experiments.