Senp1

Tissues and cells were homogenized in whole-cell lysis buffer (50 mM Tris-HCl (pH 7.6), 150 mM NaCl and 1.0% NP-40) containing a protease inhibitor cocktail and 20 mM N-ethylmaleimide to prevent SUMO de-conjugation. Immunoblotting was performed with primary antibodies against HOXA10 (1 : 1000; Santa Cruz, Santa Cruz, CA, USA), SUMO1 (1 : 1000; Abcam, Cambridge, CA, USA), SENP1 (1 : 1000; Abcam), SENP2 (1 : 1000; Abgent, San Diego, CA, USA), Flag (1 : 1000; Sigma), Myc (1 : 5000; Invitrogen, Carlsbad, CA, USA), Lamin B1 (1 : 1000; Bioworld, St Louis Park, MN, USA), HSP90B (1 : 1000; Bioworld), ubiquitin (1 : 1000, Abcam), acetylated lysine (1 : 1000; CST, Danvers, MA, USA) and GAPDH (1 : 10 000; Bioworld), followed by donkey anti-goat or goat anti-rabbit secondary antibody conjugated with HRP. Detection was performed using an enhanced chemiluminescence kit (Millipore, Billerica, USA).