Fig. 2

Loss of Usp11 increases cell proliferation, motility, and metabolism. a Growth curves of primary wild-type (Usp11^+/Y) and Usp11^-/Y MEFs. n = 3. b Colony-formation efficiency of primary Usp11^+/Y and Usp11^-/Y MEFs (left). The number of colonies per well was counted (right). n = 3. c Representative plates stained with crystal violet in transformation assays of Usp11^+/Y and Usp11^-/Y MEFs with the indicated oncogenes are shown (top). Quantification of the number of transformed foci is also shown (bottom). n = 6. d Representative images of allograft tumors of Usp11^+/Y and Usp11^-/Y MEFs transformed by E1a + Ha-ras oncogenes (top). Lysates from the tumors were subjected to IB (middle). Tumor volumes at different days were measured (bottom). n = 6, p value was determined by ANOVA (***p < 0.001). e Representative images of wound-healing assays of Usp11^+/Y and Usp11^-/Y MEFs at time points t = 0, 12, and 24 h in the presence of mitomycin C (5 μg ml⁻¹) after wound introduction (left). The percentage of wound closure at the indicated time points was determined by the ImageJ 1.46r software (right). Scale bars, 100μm. n = 3. f MMP family member gene enrichment signature from (e) by microarray. n = 3. Heatmap colors represent the relative mRNA expression as indicated in the color key. g The rates of glucose uptake, lactate production, and glutamine consumption of Usp11^+/Y and Usp11^-/Y MEFs were measured and normalized to cell number. n = 3. h The rates of glucose uptake, lactate production, and glutamine consumption of MEFs overexpressing USP11 were measured and normalized to cell number. n = 3. i Cytosolic and plasma membrane fractionation of GLUT1 in MEFs overexpressing USP11 treated with insulin (0.5 μg ml⁻¹) for the indicated times is shown. # indicates nonspecific band. Error bars represent   ±   SEM. p Value was determined by Student’s t test (*p < 0.05; **p < 0.01; ***p < 0.001). IB, immunoblotting; MEFs, mouse embryonic fibroblasts