HEK 293 cells were cultured in DMEM medium (Corning) containing fetal bovine serum (FBS; 10%, Invitrogen) and penicillin/streptomycin (PS; 1%, Thermo Fisher Scientific) on six-well tissue culture plates. Cells were manually passaged approximately every 3 d or until ∼80% confluence. Cell lines were kept in 37°C incubation at 5% CO2.

For Western blotting, protein samples were collected in Laemmli sample buffer (Thermo Fischer Scientific) and heated to 100°C for 10 min. Proteins were resolved by SDS-PAGE (Bio-Rad) and transferred to PVDF membranes using a semi-dry blotter (Bio-Rad). Membranes were blocked with 5% non-fat milk and then probed with primary antibodies rabbit anti-Sox11 (1:1000, Abcam) or anti-GAPDH (1:2000, Cell Signaling Technology) overnight at 4°C. Membranes were washed and probed with secondary antibodies conjugated to horseradish peroxidase (Millipore), and developed with the Western Blot Substrate kit (Thermo Fischer Scientific) by detecting chemiluminescence using the ChemiDoc XRS+ imaging system (Bio-Rad).

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