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19 In brief, the samples were thawed on ice, briefly vortexed, and a 200-L aliquot was transferred to a clean microcentrifuge tube

10.1096/fj.11-198416 209 SunY.SavaneninA.ReddyP

Nrf2 is controlled by two distinct beta-TrCP recognition motifs in its Neh6 domain, one of which can be modulated by GSK-3 activity

The PVDF membrane that contained the protein was blocked with 1% (w/v) skim milk in Tris-buffered saline with Tween-20 (TBST) and then incubated overnight with primary antibodies against NF-B (1:500), Akt (1:500), p-Akt (1:1000) (Abcam, UK), P-p65 (1:1000), P38 (1:1000), and p-P38 (1:800) (Cell Signaling, MA, USA) at 4C

Each treatment cycle takes 35-60 minutes, and patients usually choose to have multiple cycles performed during a single visit, usually between two and four