Western Blot Analysis of BCL11A and LRF

Western blot analysis was performed as described earlier^{27 (link)}. Briefly, whole-cell lysates were prepared using radioimmunoprecipitation assay buffer containing phenylmethanesulfonylfluoride (Sigma-Aldrich) and Halt Protease Inhibitor Cocktail (Thermo Scientific, Rockford, IL, USA). The lysate (30 μg) was loaded on a 7% sodium dodecyl sulfate-polyacrylamide gel and the western blotting was performed using the primary antibodies, anti-BCL11A (1:1000 dilution) (Cell Signaling Technologies, Danvers, MA, USA), anti-LRF (1:1000 dilution) (Cell Signaling Technologies) and anti-actin (1:5000 dilution) (BD Pharmingen), and the secondary antibodies, anti-mouse IgG HRP (Cell Signaling Technologies) and anti-rabbit IgG HRP (Invitrogen Corporation, Camarillo, CA, USA).