After compound treatment, cells were washed with PBS. Free thiols were blocked with 100 mM Pierce N-ethylmaleimide (#23030, Thermo Fisher Scientific) for 10 min. Cells were lysed by addition of 1% NP-40 and scraping. Reduced samples were mixed with NuPAGE LDS Sample Buffer (NP0008, Thermo Fisher Scientific) and 100 mM DTT and heated at 95°C for 5 min. Nonreduced samples were mixed with Pierce LDS Sample Buffer, Non-Reducing (#84788, Thermo Fisher Scientific). Protein (15 μg) was loaded onto 10% TGX gels (#4568033, Bio-Rad). Proteins were transferred to nitrocellulose membranes and blocked for 30 min in Odyssey blocking buffer 1:1 diluted with tris-buffered saline with 0.1% Tween 20 (Merck Millipore, Germany). Primary antibodies were incubated overnight at 4°C with mild agitation. Secondary antibodies were incubated for an hour at room temperature (IRDye 800CW goat anti-rabbit conjugated). Blots were imaged using an Odyssey FC Imaging System (LI-COR Biotechnology). Anti-pSTAT3 (Y705) (#D3A7; 1:1000) and anti-STAT3 (#79D7; 1:2000) were purchased from Cell Signaling Technology, anti-Prx2 (EPR5154; 1:20,000) was purchased from Abcam, and anti–glyceraldehyde-3-phosphate dehydrogenase (sc-25778; 1:1000) was purchased from Santa Cruz Biotechnology.

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