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After blocking with 5% BSA for 1 h, the membranes were incubated with anti--SMA (1:1,000 dilution, Abcam, USA) anti-Nrf2 (1:1,000 dilution, Proteintech Group, Rosemont, IL, USA), anti-HO-1 (1:1,000 dilution, Proteintech Group, Rosemont, IL, USA), anti-GPX4 (1:1,000 dilution, Abmart, Shanghai, China), anti-xCT (1:1,000 dilution, Shanghai, China), anti--tubulin antibodies (1:1,000 dilution, Cell Signaling Technology, Danvers, MA, USA) or anti-Lamin-B1 antibodies (1:10,000 dilution, Proteintech Group, Rosemont, IL, USA) at 4 C overnight, followed by horseradish peroxidase-linked anti-mouse (1:1,000 dilution, Cell Signaling Technology, Danvers, MA, USA) or anti-rabbit secondary antibodies (1:2,000 dilution, Cell Signaling Technology, Danvers, MA, USA) at 37 C for 1 h

However, there are drawbacks to using this in evaluation of NACs effectiveness on hangovers

doi: 10.3389/fmicb.2019.02324 269 RaphaelINalawadeSEagarTNForsthuberTG

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However, its mechanism of action needs to be further studied (Zhao et al., 2011)

Fallon, Nicholas, et al., Functional Connectivity with the Default Mode Network Is Altered in Fibromyalgia Patients. Plos One , vol