Overexpression of Bruton Tyrosine Kinase Inhibits the Prolif

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Western Blot. Total protein was extracted from cell lysatesafter cells were harvested and centrifuged at 4°C and 16,000 × gfor 20 min. Protein concentrations were measured by theBCA method (Beijing Dingguo Changsheng BiotechnologyCo., Ltd. company, China). Subsequently, proteins wereseparated via 12% sodium dodecyl sulfate–polyacrylamide gelelectrophore-sis for 1.5 hr and transferred onto polyvinylidenefluoride (PVDF) membranes using a wet transfer

electrophoresis tank for 1 hr. Sealed with 5% skim milk for 1hr at room tempera-ture. Wash with Tris-buffered salinewith Tween-20 (TBST) solution three times for 10 min eachtime. PVDF membranes were then incubated overnight at 4°C with the following