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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