Total protein was extracted using RIPA buffer (Sigma-Aldrich Corp.), and the protein concentrations were quantified by BCA protein assay kit (Boster Ltd., Wuhan, China). Total protein (50 μg) was separated by 10% SDS-PAGE and transferred onto polyvinylidene fluoride membranes (PVDF; Millipore, Billerica, MA, USA). The membranes were incubated with polyclonal rabbit anti-SATB1 antibody (1:2,000; Proteintech, Chicago, IL, USA), and then incubated with horseradish peroxidase-conjugated secondary antibody (1:5,000; Proteintech). The ECL-based detection was performed with enhanced chemiluminescent reagent kit (Pierce Biotechnology Inc., Rockford, IL, USA) according to the manufacturer's instructions. Protein bands were visualized using the ECL plus detection system (Pierce Biotechnology Inc.). The level of α-tubulin was used as a loading control.

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