Fig. 6

MCL cells turn on a sumoylation program prior to mitosis entry which can be targeted by TAK-981. A Jeko cell were synchronized with Palbociclib (500 nM, 24 h) followed by drug washout and treatment with nocodazole (50 ng/mL) either in the presence of DMSO or TAK-981 (100 nM) for 24 h. (left) Lysates were immunoprecipitated with antibodies directed towards SUMO1, SUMO2/3 or respective IgG controls. (right) Microscopy was performed for SUMO1 or SUMO 2/3. B (top left). Immunoprecipitants from each condition in A were analyzed by Orbitrap MS (see “Methods” section) and proteins identified as being significantly enriched (n = 3 independent experiments, p = 0.05) in SUMO IP compared to IgG are shown (left, SUMO1 = red; SUMO2/3 = blue). C Protein–protein interaction network of all sumoylation enriched proteins created by STRING using k-means clustering. D Average percent inhibition (n = 3) of sumoylated proteins with TAK-981 (100 nM) based on Orbitrap MS spectral data of individual proteins identified in B (SUMO1 substrates = red; SUMO2/3 substrates = blue)